Novel 3-acetoxymethyl-7-(iminoacetamido)-cephalosporanic acid derivatives

ABSTRACT

Compounds of the formula ##STR1## wherein R is selected from the group consisting of hydrogen and groups easily removable by acid hydrolysis of hydrogenolysis, R&#39; is selected from the group consisting of hydrogen, alkyl of 1 to 4 carbon atoms, alkenyl and alkynyl of 2 to 4 carbon atoms and groups easily removable by acid hydrolysis or hydrogenolysis. A is selected from the group consisting of hydrogen, alkali metal and equivalents of an alkaline earth metal or magnesium and an organic amine base with the proviso that when R&#39; is a group easily removable by acid hydrolysis of hydrogenolysis, R is also and when R&#39; is hydrogen, R also is hydrogen and the wavy line means the OR&#39; group may be in either one of the two possible syn or anti positions having antibiotic activity and process for their preparation.

PRIOR APPLICATION

This application is a continuation of U.S. patent application Ser. No.181,658 filed Apr. 14, 1988 now abandoned which is a division of U.S.patent application Ser. No. 449,680 filed Dec. 14, 1982 now U.S. Pat.No. 4,843,164 which is a division of U.S. patent application Ser. No.257,984 filed Apr. 27, 1981 now U.S. Pat. No. 4,376,203 which in turn isa continuation of U.S. patent application Ser. No. 71,295 filed Aug. 30,1979 now abandoned which in turn is a division of copending U.S. patentapplication Ser. No. 917,985 filed Jun. 22, 1978 now U.S. patentapplication Ser. No. 817,114 filed July 19, 1977, now U.S. Pat. No.4,152,432 which in turn is a continuation-in-part of copending commonlyassigned U.S. patent application Ser. No. 761,270 filed Jan. 21, 1977,now abandoned.

STATE OF THE ART

French patents Nos. 2,137,899, 2,137,900, 2,123,545 and 2,294,690 relateto 3-acetoxymethyl-7-amino-cephalosporanic acid compounds.

OBJECTS OF THE INVENTION

It is an object of the invention to provide the novel cephalosporanicacid derivatives of formula I and novel processes and intermediates fortheir preparation.

It is another object of the invention to provide novel antibioticcompositions and a novel method of combatting bacterial infections inwarm-blooded animals.

These and other objects and advantages of the invention will becomeobvious from the following detailed description.

THE INVENTION

The novel 3-acetoxymethyl-7-(iminoacetamido)-cephalosporanic acidderivatives of the invention have the formula ##STR2## wherein R isselected from the group consisting of hydrogen and groups easilyremovable by acid hydrolysis of hydrogenolysis, R' is selected from thegroup consisting of hydrogen, alkyl of 1 to 4 carbon atoms, alkenyl andalkynyl of 2 to 4 carbon atoms and groups easily removable by acidhydrolysis or hydrogenolysis, A is selected from the group consisting ofhydrogen, alkali metal and equivalents of an alkaline earth metal ormagnesium and an organic amine base with the proviso that when R' is agroup easily removable by acid hydrolysis or hydrogenolysis, R is alsoand when R' is hydrogen, R also is hydrogen and the wavy line means theOR' group may be in either one of the two possible syn or antipositions.

The groups easily removable by acid hydrolysis or by hydrogenolysis arewell known in cephalosporin chemistry and examples thereof aretert.-butoxycarbonyl, trityl, benzydibenzyl, trichloroethyl,carbobenzyloxy, formyl, trichloroethoxycarbonyl or 2-tetrahydropyranyl.R' may also be alkyl, alkenyl or alkynyl such as methyl, ethyl, propyl,isopropyl, butyl, sec.-butyl, tert.-butyl, vinyl, propenyl, butenyl,ethynyl or propargyl.

Examples of A are hydrogen, potassium, sodium, lithium, calcium,magnesium, trimethylamine, triethylamine, methylamine, propylamine,N,N-dimethylethanolamine, tris-(hydroxymethyl)-aminomethane, arginine orlysine.

The groups easily removable by acid hydrolysis or hydrogenolysis thatare preferred are tert.-butoxycarbonyl, trityl, dibenzyl, trichloroethyland carbobenzyloxy.

The compounds of formula I may exist in the form of the syn isomer ofthe formula ##STR3## or in the form of the anti isomer of the formula##STR4## but the OR' group is preferably in the syn position.

Preferably, in the compounds of formula I, R is hydrogen or trityl, R'is hydrogen, trityl, alkyl of 1 to 4 carbon atoms or alkenyl or alkynylof 2 to 4 carbon atoms and A is hydrogen, sodium or diethylamine.

Preferred compounds of formula I are3-acetoxymethyl-7-[2-(2-amino-4-thiazolyl)-2-methoxyiminoacetamido]-ceph-3-eme-4-carboxylicacid, sodium3-acetoxymethyl-7-[2-(2-amino-4-thiazolyl)-2-methoxyiminoacetamido]-ceph-3-4-carboxylate,syn isomer of3-acetoxymethyl-7-[2-(2-amino-4-thiazolyl)-2-methoxyiminoacetamido]-ceph-3-eme-4-carboxylicacid, syn isomer of sodium3-acetoxymethyl-7-[2-(2-amino-4-thiazolyl)-2-methoxyiminoacetamido]-ceph-3-eme-4-carboxylate,3-acetoxymethyl-7-[2-(2-amino-4-thiazolyl)-2-methoxyiminoacetamido]-ceph-3-eme-4-carboxylic acid as obtained by the process described in Example4, sodium3-acetoxymethyl-7-[2-(2-amino-4-thiazolyl)-2-methoxyiminoacetamido]-ceph-3-eme-4-carboxylateas obtained by the process described in Example 7, syn isomer ofcrystalline sodium3-acetoxymethyl-7-[2-(2-amino-4-thiazolyl)-2-methoxyiminoacetamido]-ceph-3-eme-4-carboxylate,syn isomer of3-acetoxymethyl-7-[2-(2-amino-4-thiazolyl)-2-ethoxyiminoacetamido]-ceph-3-eme-4-carboxylicacid and its alkali metal, alkaline earth metal, magnesium and organicamine salts syn isomer of 3-acetoxymethyl-7-[2-(2-amino-4-thiazolyl)-2-(2-propenyloxyimino)-acetamido]-ceph-3-eme-4-carboxylic acid and itsalkali metal, alkaline earth metal, magnesium and organic amine salts,syn isomer of3-acetoxymethyl-7-(2-(2-amino-4-thiazolyl)-2-(1-methylethoxyimino)-acetamido)ceph-3-eme-4-carboxylicacid and its alkali metal, alkaline earth metal, magnesium and organicamine salts and syn isomer of7-(2-(2-amino-4-thiazolyl)-2-hydroxyiminoacetamido)-3-acetoxymethyl-ceph-3-eme-4-carboxylic acid and their alkali metal, alkaline earth metal, magnesiumand organic amine salts especially the sodium salt and syn isomer of7-(2-(2-trityl amino-4-thiazolyl)-2-tritylhydroxyiminoacetamido)-3-acetoxymethyl-ceph-3-eme-4-carboxylic acid.

The products of the invention may exist in the form indicated in formulaI or in the form of products of the formula ##STR5##

The process of the invention for the preparation of the compounds offormula I comprises reacting 7-amino-cephalosporanic acid with an acidof the formula ##STR6## or a derivative thereof wherein R₁ is a groupeasily removable by acid hydrolysis or hydrogenolysis and R₁ is selectedfrom the group consisting of alkyl of 1 to 4 carbon atoms, alkenyl andalkynyl of 2 to 4 carbon atoms and a group easily removable by acidhydrolysis or hydrogenolysis to form a compound of the formula ##STR7##and the latter may be subjected to acid hydrolysis or hydrogenolysis toobtain a compound of formula I wherein R and A are hydrogen and R' ishydrogen, alkyl of 1 to 4 carbon atoms or alkenyl or alkynyl of 2 to 4carbon atoms and the said product Ia and I may be satisfied by knownmethods.

In a preferred mode of the said process, 7-aminocephalosporanic acid isreacted with a functional derivative of the acid of formula II such asthe acid chloride or anhydride such as that formed in situ with isobutylchloroformate or dicyclohexylcarbodiimide with the acid. Other acidhalides other acid anhydrides formed in situ with other alkylchloroformates, a dialkylcarbodiimide or other dicycloalkylcarbodiimidemay be used. Equally useful are other acid derivatives such as the acidazide, active amide or ester of the acid such as formed byhydroxysuccimide, p-nitrophenol or 2,4-dinitrophenol.

The reaction of 7-amino-cephalosporanic acid with isobutyl chloroformateanhydride or acid chloride of the acid of formula II is preferablyeffected in the presence of a basic agent such as an alkali metalcarbonate or an organic tertiary amine such as N-methyl-morpholine orpyridine or a trialkylamine such as triethylamine.

The acid hydrolysis may be effected with formic acid, trifluoroaceticacid or acetic acid and may be anhydrous or in aqueous solution. Alsouseful in zinc-acetic acid system. To eliminate tert.-butoxycarbonyl ortrityl groups, the acid agent is preferably anhydrous trifluoroaceticacid or aqueous acetic acid or formic acid. The zinc-acetic acid systemis preferably used to remove a trichloroethyl group. The benzyl,dibenzyl and carbobenzyloxy groups are preferably removed byhydrogenolysis in the presence of a hydrogenation catalyst.

The salification may be effected by known methods such as by reactingthe free acid with a mineral base such as sodium hydroxide, potassiumhydroxide or sodium bicarbonate or a salt of a substituted ornon-substituted aliphatic carboxylic acid such as diethylacetic acid,ethylhexanoic acid or especially acetic acid with the preferred saltsbeing the sodium salts. The salification may be effected with an organicbase such as triethylamine.

For the preparation of the salts, the solvates of the free acids mayalso be used in place of the free acids. The solvates may be obtainedwith water, formic acid or an alcohol, for example. The solvates with analcohol, preferably ethanol, may be obtained by treatment with analcohol-water mixture of the solvents with formic acid which is thenfollowed by concentration of the solution.

The salification is preferably effected in at least one solvent such aswater, ether, methanol, ethanol or acetone and the salts may be eithercrystalline or amorphous depending on the reaction conditions used. Thecrystalline salts are preferably prepared by reacting the free acid or asolvate thereof, formed for example, with formic acid or ethanol, with asalt of the above mentioned aliphatic carboxylic acid, preferably sodiumacetate. In the preparation of a sodium salt, the reaction is effectedin the appropriate organic solvent such as methanol which can containsmall amounts of water. It is also possible to change the amorphoussalts into crystalline salts. To this effect, an amorphous sodium saltwhich can be in the form of a solvate with, for example, 0.5, 1 or 1.5moles of water is dissolved in an appropriate organic solvent,preferably a low molecular weight alcohol such as methanol.Crystallization may then be directly effected or by addition of othersolvents such as ethanol, isopropanol, n-butanol, acetone, ethers andgenerally organic solvents miscible with methanol. If the startingmaterial or the solvent or the two constituents contain water, the saltmay crystallize in the form of a hydrate. For example, the syn isomer ofsodium3-acetoxymethyl-7-[2-(2-amino-4-thiazolyl)-2-methoxyiminoacetamido]-ceph-3-eme-4-carboxylatehas been isolated with 0.5, 1 or 1.5 moles of water.

In a modification of the process of the invention to prepare a compoundof the formula ##STR8## wherein R₁ " is selected from the groupconsisting of hydrogen, alkyl of 1 to 4 carbon atoms and alkenyl andalkynyl of 2 to 4 carbon atoms, a compound of the formula ##STR9##wherein A₁ is a group easily removable by acid hydrolysis orhydrogenolysis is reacted with an acid of formula II or a functionalderivative thereof wherein R₁ ' is chloroacetyl or a group easilyremovable by acid hydrolysis or hydrogenolysis or alkyl of 1 to 4 carbonatoms or alkenyl or alkynyl of 2 to 4 carbon atoms and R₁ ischloroacetyl or a group easily removable by acid hydrolysis orhydrogenolysis to obtain a compound of the formula ##STR10## which isthen treated with one or more acid hydrolysis agents or one or morehydrogenolysis agents or with thiourea or with one or more of thesedifferent agents to recover a compound of formula Ib.

Examples of groups easily removable by acid hydrolysis or hydrogenolysisfor R₁ and R₁ ' are tert.-butoxycarbonyl, trityl, benzyl, dibenzyl,trichloroethyl, carbobenzyloxy, or formyl but also useful are2-tetrahydropyranyl and trichloroethoxycarbonyl. For A₁, the groups arepreferably benzhydryl, tert.-butyl, benzyl, p-methoxybenzyl ortrichloroethyl.

In a preferred mode of this process, a compound of formula Ic is reactedwith a functional derivative of the acid of formula II such as the acidchloride or acid anhydride such as that formed in situ with isobutylchloroformate or dicyclohexylcarbodiimide. Equally useful are other acidhalides or other mixed anhydrides formed in situ with other alkylchloroformates or other dicycloalkylcarbodiimides or adialkylcarbodiimide. Equally useful are other acid derivative such asacid azide, active amides of the acid or active esters of the acid suchas that formed with hydroxy succinimide, p-nitrophenol or2,4-dinitrophenol. When the acid halide or acid anhydride with isobutylchloroformate are used, it is preferred to effect the reaction in thepresence of a basic agent such as an alkali metal carbonate or atertiary organic amine such as N-methyl morpholine, pyridine or atrialkylamine such as triethylamine.

The compounds of formula XI are changed into compounds of formula Ibwherein R₁ and A₁ are substituents replaced by hydrogen and when R₁ ' isa group easily removed by acid hydrolysis or hydrogenolysis or ischloroacetyl, the compounds of formula XI are changed to compounds offormula Ib when R₁ ' is replaced by hydrogen. To accomplish this, thecompound of formula XI is treated with one or more acid hydrolysisagents when R₁ and A₁ are a group easily removed by acid hydrolysis andR₁ ' is a group easily removed by acid hydrolysis or alkyl or with oneor more hydrogenolysis agents when R₁ and A₁ are groups easily removedby hydrogenolysis and R'₁ is a group easily removed by hydrogenolysis oralkyl. The compounds of formula XI are treated with one or more acidhydrolysis agents and one or more hydrogenolysis agents when at leastone of R₁, A₁ and R'₁ is a group easily removed by acid hydrolysis andat least one of these groups is a group easily removed byhydrogenolysis. Finally, the compound of formula XI is treated withthiourea and optionally with one or more acid hydrolysis agents ofhydrogenolysis agents, when at least one of R₁ and R'₁ is chloroacetyl.

The acid hydrolysis may be effected with formic acid, trifluoroaceticacid or acetic acid and may be anhydrous or in aqueous solution. Alsouseful is zinc-acetic acid system. To eliminate tert.-butoxycarbonyl oftrityl groups for R₁ or R'₁ or benzhydryl, tert.-butyl or paramethoxybenzyl for A₁, the acid agent is preferably anhydrous trifluoroaceticacid or aqueous acetic acid or formic acid. The zinc-acetic acid systemis preferably used to remove a trichloroethyl group for R₁, R'₁ and A₁.The benzyl, dibenzy and carbobenzyloxy groups for R₁ and R'₁ and benzylfor R₁, R'₁ and A₁ are preferably removed by hydrogenolysis such as withhydrogen in the presence of a hydrogenation catalyst. The reaction ofthiourea with a compound of formula XI containing at least onechloroacetyl is preferably effected in a neutral or acid media asdescribed by Masaki (JACS, Vol. 90 (1964), p. 4508).

Another aspect of the invention includes the process for the preparationof compounds of formula II by reacting thiourea with a compound of theformula ##STR11## wherein R' has the above definition and AlK is alkylof 1 to 4 carbon atoms and treating the resulting product with a base toform a compound of the formula ##STR12## reacting the latter with afunctional derivative of a group easily removable by acid hydrolysis orhydrogenolysis to obtain a compound of the formula ##STR13## wherein R₁and R₁ ' have the above definitions and treating the latter with a baseand then an acid to obtain the corresponding compound of formula II.

Preferably, the base used to treat the compound of formula IV ispotassium acetate by other bases such as alkali metal carbonates orbicarbonates or dilute sodium hydroxide or potassium hydroxide are alsouseful. The functional derivative of a group which is easily removableby acid hydrolysis or hydrogenolysis is preferably trityl chloride inthe presence of triethylamine or other tertiary amine bases such aspyridine, N-methyl-morpholine or other trialkylamines. Equally usefulare tert.-butyl chloroformate formed in situ, tert.-butyl azido formate,trichloroethyl chloroformate, benzyl chloroformate, mixed formyl-aceticacid anhydride prepared in situ, benzyl and dibenzyl halides such as thechloride, phthalic anhydride or N-carbethoxyphthalimide.

The base used to treat the compound of formula V is preferably sodiumhydroxide but other bases such as potassium hydroxide or bariumhydroxide are also useful. The acid used to obtain the compound offormula II is preferably dilute hydrochloric acid but other acids suchas acetic acid or formic acid may also be used.

In a variation of the process to produce compounds of formula II, acompound of formula IV is treated first with a base and then with anacid to obtain a compound of the formula ##STR14## and reacting thelatter with a functional derivative of R₁ wherein R₁ is chloroacetyl ora group easily removably by acid hydrolysis or hydrogenolysis to obtainthe corresponding compound of formula II.

The base used for the saponification is preferably sodium hydroxide butequally useful are potassium hydroxide and barium hydroxide. Thepreferred acid is dilute hydrochloric acid but also useful are aceticacid or formic acid. The functional derivative of R₁ is preferablytrityl chloride used in the presence of triethylamine or other tertiaryamine bases such as other trialkylamines, pyridine orN-methylmorpholine. Equally useful are other easily removable groupsfunctional derivatives such as tert.-butyl chloroformate formed in situor tert.-butyl azidoformate, trichloroethyl chloroformate or benzylchloroformate, or a mixed formylacetic acid anhydride prepared in situ,benzyl or dibenzyl chloride or other halide, phthalic anhydride orN-carbethoxyphthalimide. The functional derivative of chloroacetyl ispreferably chloroacetic acid anhydride or a halide such asmonochloroacetyl chloride and in the latter case, a base as definedabove is preferably present.

Another feature of the invention is a process for the preparation of acompound of the formula ##STR15## wherein R₁ and R_(c) " are chloracetylor a group easily removed by acid hydrolysis or hydrogenolysis withoutbeing the same and Alk is alkyl of 1 to 4 carbon atoms comprisingreacting a compound of the formula ##STR16## with a functionalderivative of R_(c) ". R₁ is preferably trityl and the functionalderivative of R_(c) " is preferably dihydropyranyl to form thetetrahydropyranyl compound.

In a variation of the process of the invention to prepare compounds offormula I wherein A and R are hydrogen and R' is hydrogen, alkyl of 1 to4 carbon atoms or alkenyl or alkynyl or 2 to 4 carbon atoms, a salt of acompound of formula Ia wherein R₁ is a group easily removable by acidhydrolysis and R₁ ' is selected from the group consisting of alkyl of 1to 4 carbon atoms, alkenyl or alkynyl of 2 to 4 carbon atoms and a groupeasily removable by acid hydrolysis is treated with an acid to obtainthe desired compound or a salt of a compound of formula Ia wherein R₁ isa group easily removable by hydrogenolysis and R₁ ' is selected from thegroup consisting of alkyl of 1 to 4 carbon atoms, alkenyl or alkynyl of2 to 4 carbon atoms and a group easily removable by hydrogenolysis issubjected to hydrogenolysis to obtain the desired compound in the formof a salt. The preferred acid is formic acid but also useful aretrifluoroacetic acid or acetic acid in anhydrous or aqueous solutionform. The hydrogenolysis is preferably effected with a hydrogenationcatalyst.

A variation of the process to prepare a compound of the formula##STR17## wherein AlK is alkyl of 1 to 4 carbon atoms and R_(a) ' isalkyl of 1 to 4 carbon atoms comprises reacting a compound of theformula ##STR18## with an alkylation agent to obtain a compound of theformula ##STR19## reacting the latter with a bromination agent to form acompound of the formula ##STR20## and reacting the latter with thioureaand then a base to obtain the compound of formula IVa.

The alkylation agent used to treat the compound of formula VI ispreferably an alkyl halide such as an alkyl chloride, bromide or iodideor an alkyl sulfate of 1 to 4 alkyl carbon atoms. The bromination agentreacted with the compound of formula VII is preferably bromine and thebase used to obtain the product of formula IVa is preferably an alkalimetal carbonate or bicarbonate but equally useful are dilute sodium orpotassium hydroxide or potassium acetate.

A variation of the process to produce the compounds of formula I whereinA and R are hydrogen and R' is hydrogen, alkyl of 1 to 4 carbon atoms oralkenyl or alkynyl of 2 to 4 carbon atoms comprises reacting7-amino-cephalosporanic acid with a compound of the formula ##STR21##wherein R_(a) " is selected from the group consisting of chloroacetyl,alkyl of 1 to 4 carbon atoms and alkenyl and alkynyl of 2 to 4 carbonatoms or a functional derivative thereof to obtain a compound of theformula ##STR22## and reacting the latter with thiourea to obtain thecorresponding compound of formula I.

The reaction with 7-amino-cephalosporanic acid is effected under thesame conditions as the reaction with a compound of formula II. Thereaction with thiourea is preferably effected in a neutral or acid mediawith the type of reaction described by Masaki [J.A.C.S., Vol. 90 (1968),p. 4508].

A process for the preparation of a compound of formula IIa comprisesreacting a functional derivative of chloroacetic acid with a compound ofthe formula ##STR23## wherein R₁ " is hydrogen, alkyl of 1 to 4 carbonatoms of alkenyl or alkynyl of 2 to 4 carbon atoms and alk is alkyl of 1to 4 carbon atoms to obtain a compound of the formula ##STR24## whereinR_(a) " has the above definition and treating the latter with first abase and then an acid to obtain a compound of formula IIa.

The functional derivative of chloroacetic acid is preferablychloroacetic acid anhydride or chloroacetyl halide such as the chloride.If the acid halide is used, the reaction is preferably effected in thepresence of a basic agent as discussed previously. The base used totreat the compound of formula Va is preferably sodium hydroxide butother bases such as potassium hydroxide or barium hydroxide may be used.The acid used to obtain a compound of formula IIa is preferably dilutehydrochloric acid but equally useful are acetic acid or formic acid.

A variant of the process to obtain a compound of the formula ##STR25##wherein R₁ and AlK have the above definition and R_(b) " is alkyl of 1to 4 carbon atoms or alkenyl or alkynyl of 2 to 4 carbon atoms comprisestreating a compound of the formula ##STR26## with a functionalderivative of a group easily removable by acid hydrolysis orhydrogenolysis to obtain a compound of the formula ##STR27## andreacting the latter with an alkylation agent to obtain the compound offormula Vb.

The said functional derivative is preferably trityl chloride used in thepresence of a base preferably triethylamine although other bases such asN-methyl-morpholine, pyridine or other trialkylamine may be used. Otheruseful functional derivatives are tert.-butyl chloroformate, tert.-butylazidoformate, trichloroethyl chloroformate, dibenzyl chloroformate,mixed formyl-acetic acid anhydride formed in situ, benzyl halides anddibenzyl halides such as the chloride, phthalic anhydride orN-carbethoxyphthalimide. The alkylation agent is preferably an alkylsulfate or an alkyl halide such as the iodide.

The syn and anti configuration of the products of formula I isdetermined by the configuration of the products of formula IV since thelatter configuration is maintained in the synthesis. The same is truefor the products of formula IVa, IVb and IV" as they are within thescope of formula IV. The configuration of the products of formula IVdepends upon a certain number of parameters of the process to preparethe products.

It can also be stated that the action of thiourea with the products offormula III is effected either in an aqueous solvent such as aqueousacetone or aqueous ethanol or at room temperature with a substantiallystoichiometric amount of thiourea stirred therewith for a reaction timeof 1 to 3 hours or the said conditions can be combined to obtain the synisomer.

The process of the invention for the preparation of compounds of formulaIV in the syn form comprises reacting thiourea with a compound of theformula ##STR28## wherein X is chlorine or bromine, AlK is alkyl of 1 to4 carbon atoms and R_(b) ^(') is selected from the group consisting ofhydrogen, alkyl of 1 to 4 carbon atoms, alkenyl and alkynyl of 2 to 4carbon atoms and a group easily removable by acid hydrolysis orhydrogenolysis when X is chlorine or hydrogen or alkyl of 1 to 4 carbonatoms when X is bromine either in an aqueous solvent, or at roomtemperature with a substantially stoichiometric amount of thiourea for afew hours, or by a combination of the said conditions.

The antibiotic compositions of the invention are comprised of anantibiotically effective amount of at least one compound of formula Iand an inert pharmaceutical carrier. The compositions may be in the formof tablets, dragees, gelules, granules, suppositories, injectablesolutions or suspensions, creams, pomades, gels, etc. prepared in theusual fashion.

Examples of suitable excipients or pharmaceutical carriers are talc,arabic gum, latose, starch, magnesium stearate, cacao butter, aqueous ornon-aqueous vehicles, fatty bodies of vegetable or animal origin,paraffinic derivatives, glycols, preservatives, diverse wetting agents,dispersants and emulsifiers. The compositions of the invention andparticularly those containing the compounds of formula I in the syn formpossess very good antibiotic activity against gram positive bacteriasuch as staphylococcus, streptococcus, particularly penicilliniresistant staphylococcus as well as against gram negative bacteria suchas coliform bacteria, Klebsiella, Salmonella and Proteus.

The compositions are therefore useful in the treatment of germ sensitiveinfections and particularly those of staphylococcia such asstaphylococcal septicemia, staphylococcia malignant on the face or skin,pyodermatitis, septic or suppurantes sores, anthrax, phlegmons,eresipels, acute primitive or post-grip staphylococcia, bronchopneumoniaor pulmonary suppurations. They are equally useful for the treatment ofcollibacillosis and associated infections, infections of Proteus,Klebsiella and Salmonella and other infections caused by gram negativebacteria. Among the preferred compositions are those containing thecompounds of formula I with the syn form. Preferably R is hydrogen, R'is hydrogen, alkyl of 1 to 4 carbon atoms and alkenyl and alkynyl of 2to 4 carbon atoms and A is hydrogen and sodium.

Among the preferred compositions are also those containing as activeprinciple the following products:

3-acetoxymethyl 7-[2-(2-amino 4-thiazolyl) 2-methoxy imino acetamido]ceph-3-eme 4-carboxylic acid, syn isomer, this product is especiallydescribed in example 4, 6, 20 pr 22,

the sodium salt of 3-acetoxymethyl 7-[2-(2-amino 4-thiazolyl) 2-methoxyimino acetamido] ceph-3-eme 4-carboxylic acid, syn isomer, this productis especially described in example 7,

the crystalline sodium salt of 3-acetoxymethyl7-[2-(2-amino-4-thiazoly)-2-methoxyiminoacetamido]-ceph-3-eme-4-carboxylicacid, syn isomer, or the following products:

3-acetoxymethyl-7[2-(2-amino-4-thiazolyl)-2-ethoxyiminoacetamido]-ceph-3-eme-4-carboxylicacid, syn isomer and its pharmaceutically acceptable salts with analkali metals, the alkaline-earth metals, magnesium or the organic aminobase,

3-acetoxymethyl-7-[2-(2-amino-4-thiazolyl)-2-((2-propenyl)-oxyimino)-acetamido]-ceph-3-eme-4-carboxylicacid, syn isomer and its pharmaceutically acceptable salts with thealkali metals, the alkaline earth metals, magnesium or the organic aminobases,

3-acetoxymethyl-7-[2-(2-amino-4-thiazolyl)-2((1-methylethoxy)-imino)-acetamido]-ceph-3-eme-4-carboxylicacid, syn isomer and its pharmaceutically acceptable salts with thealkali metals, the alkaline earth metals, magnesium or the organic aminobases,

syn isomer of7-[2-(2-amino-4-thiazolyl)-2-hydroxyimino-acetamido]-3-acetoxymethyl-ceph-3-eme-4-carboxylicacid and its non-toxic, pharmaceutically acceptable salts with alkalimetals, alkaline earth metals, magnesium and organic amine bases andespecially the free acid and its sodium salt.

The novel method of the invention for combatting bacterial infections inwarm-blooded animals, including humans, comprises administering towarm-blooded animals an antibacterially effective amount of at least onecompound of formula I. The compounds may be administered orally,rectally, parenterally or locally by topical application to the skin ormucous. The usual daily dose is 5 to 80 mg/kg depending on the compoundand the method of administration.

Among the novel intermediate compounds of the invention are compounds ofthe formula ##STR29## wherein R₂ is selected from the group consistingof hydrogen and a group easily removable by acid hydrolysis orhydrogenolysis and R₂ ^(') is selected from the group consisting ofhydrogen, alkyl of 1 to 4 carbon atoms, alkenyl and alkynyl of 2 to 4carbon atoms and a group easily removable by acid hydrolysis orhydrogenolysis with the proviso that R₂ ^(') is not hydrogen when R₂ isother than hydrogen and the novel compounds of the formula ##STR30##wherein AlK is alkyl of 1 to 4 carbon atoms and R₁ ^(') is selected fromthe group consisting of alkyl or 1 to 4 carbon atoms, alkenyl andalkynyl of 2 to 4 carbon atoms and a group easily removable by acidhydrolysis or hydrogenolysis and the novel compounds of the formula##STR31## wherein R_(a) ^(") is selected from the group consisting ofchloroacetyl, alkyl of 1 to 4 carbon atoms and alkenyl and alkynyl of 2to 4 carbon atoms as well as compounds of the formula ##STR32## whereinR_(I), R₁ ^(') and A₁ have the above definitions. The latter twoproducts also possess antibiotic properties similar to those of formulaI and may be used in the same manner.

The compounds of formula III that are not known may be preparedbeginning with ethyl γ-chloro-α-oximino-acetylacetate described in J. ofMedicinal Chemistry, Vol. 16, 1973, No. 9. To obtain compounds where R'is alkyl of 1 to 4 carbon atoms or alkenyl or alkynyl of 2 to 4 carbonatoms, the said ethyl ester is reacted with the correspondinghydrocarbon halide or sulfate. To obtain compounds wherein R' is a groupeasily removable by acid hydrolysis or hydrogenolysis the said ester isreacted by classical reactions with functional derivatives of saidgroups.

The products of formula III' wherein X is bromine and R_(b) ^(') ishydrogen may be prepared by reacting a compound of the formula ##STR33##with a bromination agent under the same conditions described for thetreatment of compounds of formula VII.

In addition to the compounds prepared in the specific examples, thefollowing compounds in the syn isomer form are also part of thepreferred group:3-acetoxymethyl-7-[2-(2-amino-4-thiazolyl)-2-propyloxyiminoacetamido]-ceph-3-eme-4-carboxylicacid,3-acetoxymethyl-7-[2-(2-amino-4-thiazolyl)-2-butyloxyiminoacetamido]-ceph-3-eme-4-carboxylicacid,3-acetoxymethyl-7-[2-(2-amino-4-thiazolyl)-2-(2-methylpropyloxyimino)-acetamido]-ceph-3-eme-4-carboxylicacid,3-acetoxymethyl-7-[2-(2-amino-4-thiazolyl)-2-(1,1-dimethylethoxyimino)-acetamido]-ceph-3-eme-4-carboxylicacid,3-acetoxymethyl-7[2-(2-amino-4-thiazolyl)-2-(2-butenyloxyimino-acetamido]-ceph-3eme-4-carboxylicacid,3-acetoxymethyl-7-[2-(2-amino-4-thiazolyl)-2-(3-butenyloxyimino-acetamido]-ceph-3-eme-4-carboxylicacid,3-acetoxymethyl-7[2-(2-amino-4-thiazolyl)-2-(2-butenyloxyimino)-acetamido]-ceph-3-eme-4-carboxylicacid,3-acetoxymethyl-7-[2-(2-amino-4-thiazolyl)-2-(3-butenyloxyimino)-acetamido]-ceph-3-eme-4-carboxylicacid,3-acetoxymethyl-7-[2-(2-amino-4-thiazolyl)-2-vinyloxyiminoacetamido]-ceph-]3-eme-4-carboxylicacid,3-acetoxymethyl-7-[2-(2-amino-4-thiazolyl)-2-(1-propenyloxyimino)-acetamido]-ceph-3-eme-4-carboxylicacid,3-acetoxymethyl-7-[2-(2-amino-4-thiazolyl)-2-(1-propynyloxyimino)-acetamido]-ceph-3-eme-4-carboxylicacid,3-acetoxymethyl-7-]2-(2-amino-4-thiazolyl)-2-(1-butenyloxyimino)-acetamido]-ceph-3-eme-4-carboxylicacid,3-acetoxymethyl-7-[2-(2-amino-4-thiazoly)-2-(1-ethynyloxyimino)-acetamido]-ceph-3-eme-4-carboxylicacid, 3-acetoxymethyl-7-[2-(2-amino-4thiazolyl)-2-(2-propynyloxyimino)-acetamido]-ceph-3-eme-4-carboxylicacid and3-acetoxymethyl-7-[2-(2-amino-4-thiazolyl)-2-(1-butenyloxyimino)-acetamido]-ceph-3-eme-4-carboxylicacid and their salts with alkali metals, alkaline earth metals,magnesium and organic amines.

In the following examples there are described several preferredembodiments to illustrate the invention. However, it should beunderstood that the invention is not intended to be limited to thespecific embodiments.

EXAMPLE 13-acetoxymethyl-7-[2-(2-tritylamino-4-thiazolyl)-2-tritylhydroxyiminoacetamido]-ceph-3-eme-4-carboxylicacid STEP A: ethyl 2-(2-amino-4-thiazolyl)-2-hydroxyiminoacetate

A mixture of 2 g of ethyl γ-chloro-α-oximinoacetylacetate, 5 ml ofethanol and 0.76 g of thiourea was stirred at room temperature for 16hours while the hydrochloride crystallized and after dilution with 5 mlof ether, the mixture was vacuum filtered. The filter was rinsed with a1-1 ethanol-ether mixture and then ether to obtain 1.55 g of thehydrochloride salt which was dissolved at 40°-50° C. in 8 ml of water.The solution was neutralized to a pH of 5-6 by addition of sodiumacetate and the free ethyl 2-(2-amino-4-thiazolyl)-2-hydroxyiminoacetatecrystallized. The mixture was iced and then was vacuum filtered and theproduct was washed with water and dried to obtain 1.22 g of theantiisomer melting at 154° C.

The combined wash waters were concentrated and the residue was taken upin water. The solution was washed with ether and sodium bicarbonate wasadded thereto. The mixture was vacuum filtered and the product waswashed with water to obtain 1.9 g of a product having two spots in thinlayer chromatography. The product was chromatographed over silica gel,was eluted with ether and the combined pure fractions of theconcentrated syn isomer were empasted with ether, vacuum filtered anddried to obtain 50 mg of the syn isomer.

STEP B: ethyl 2-(2-tritylamino-4-thiazolyl)-2-tritylhydroxyiminoacetate

A solution of 15 g of trityl chloride in 30 ml of chloroform was addedat 10° C. to a solution of 5.4 g of the product of Step A in 54 ml ofchloroform and 7.5 ml of triethylamine and after standing for an hour,the mixture was washed with 40 ml of water and then 20 ml of watercontaining 4 ml of N hydrochloric acid. The mixture was decanted and theorganic phase was dried and evaporated to dryness. The residue was takenup in 10 ml of ether and 50 ml of methanol were added with stirring. Themixture was vacuum filtered and the product was washed with methanol toobtain in 2 crops. 14.2 g of ethyl2-(2-tritylamino-4-thiazolyl)-2-tritylhydroxyiminoacetate.

STEP C: 2-(2-tritylamino-4-thiazolyl)-2-tritylhydroxyiminoacetic acid

A suspension of 10.5 g of the ester of Step B in 55 ml of dioxane wasbrought to approximately reflux and 17 ml of 2N sodium hydroxide wereslowly added thereto. The mixture was brought to a slight reflux and wascooled and vacuum filtered. The salt product was taken up in 60 ml ofmethylene chloride, 20 ml of water and 2 ml of acetic acid and themixture was vacuum filtered. The product was washed with water to obtaina first crop of 7 g of2-(2-tritylamino-4-thiazolyl)-2-trithylhydroxyimino-acetic acid. Thedioxane was evaporated from the filtrate and 20 ml of methylenechloride, 10 ml of water and 1 ml of acetic acid were added to obtain asecond crop of 1.5 g of the said acid for a total yield of 8.5 g.

Analysis: C₄₃ H₃₃ O₃ N₃ S . 0.5 H₂ O

Calculated: %C, 75.85,H, 5.03%N, 6.17%S, 4.7.

Found: %C, 75.8,%H, 4.9%N, 5.9,%S, 4.6.

STEP D:3-acetoxymethyl-7-[2-(2-tritylamino-4-thiazolyl)-2-tritylhydroxyiminoacetamido]-ceph-3-eme-4-carboxylicacid

5 ml of N-methyl-morpholine were added with stirring to a suspension of8.5 g of the acid of Step C and 50 ml of methanol and after stirring themixture for 10 minutes at 30° C., 30 ml of methylene chloride were addedthereto. The mixture was concentrated and 100 ml of ether were addedthereto. The mixture effloresced and was vacuum filtered. The recoveredprecipitate was washed with ether and dried to obtain a first crop ob7.2 g of the N-methyl-morphine salt. The filtrate was evaporated todryness and the residue was taken up in ether to obtain a second crop ofthe same salt.

A suspension of 4.24 g of the salt in 60 ml of methylene chloride wasstirred under an inert gas for 5 minutes and after cooling to -5° C., 6ml of a molar solution of isobutyl chloroformate in methylene chloridewere added thereto. The mixture was stirred at -5° C. for 15 minutes andwas then cooled to -20° C. after which a solution of 1.36 g of7-amino-cephalosporanic acid in 25 ml of methylene chloride and 1.4 mlof triethylamine was added. The mixture stood for an hour at roomtemperature and was washed with 50 ml of water containing 10 ml ofN-hydrochloric acid and was vacuum filtered. The filtrate was decantedand the organic phase was washed with water and evaporated to dryness.The residue was triturated with ether, vacuum filtered and washed withether to obtain 4.5 g of raw product. The raw product in 10 ml ofmethylene chloride was stirred for an hour at 10° C. and was vacuumfiltered. The filter was rinsed with methylene chloride and 50 ml ofether were added to the filtrate. The mixture was stirred and vacuumfiltered and the precipitate was washed with ether to obtain 2.29 g of3-acetoxymethyl-7-[2-(2-tritylamino-4-thiazolyl)-2-tritylhydroxyiminoacetamido]-ceph-3-eme-4-carboxylicacid. A second crop of 0.856 g of the said product was also obtained fora total yield of 3.146 g.

EXAMPLE 23-acetoxymethyl-7-[2-(2-amino-4-thiazolyl)-2-hydroxyiminoacetamido]ceph-3-eme-4-carboxylicacid

A suspension of 2.29 g of the acid of Example 1 in 18.4 ml of 50%aqueous formic acid was stirred at 55° C. for 15 minutes and was thencooled. 10 ml of water were added thereto and the mixture was vacuumfiltered. The filtrate was washed with water and concentrated to drynessunder reduced pressure. The residue was added to acetone and the mixturewas vacuum filtered. 30 ml of ether were added to the filtrate and themixture was stirred and vacuum filtered. The recovered produce waswashed with ether to obtain 0.665 g of3-acetoxymethyl-7-[2-(2-amino-4-thiazolyl)-2-hydroxyiminoacetamido]-ceph-3-eme-4-carboxylicacid. A second crop of 0.123 g of the product was obtained bycrystallization for a total yield of 0.788 g. 70 mg of carbon black wereadded to a solution of 0.735 g of the said product in 7.5 ml of ethanoland 7.5 ml of acetone and the mixture was vacuum filtered. The filtratewas evaporated to dryness and the residue was effloresced with ethanoland washed with ethanol to obtain a first yield of 0.450 g and a secondyield of 0.105 g of3-acetoxymethyl-7-[2-(2-amino-4-thiazolyl)-2-hydroxyiminoacetamido]-ceph-3-eme-4-carboxylicacid.

Infrared Spectrum:

Carbonyl at 1774, 1740 and 1676^(cm-1) ; --C═C--NH₂ at 1630^(cm-1) andNH at 1520^(cm-1)

EXAMPLE 33-acetoxymethyl-7-[2-(2-tritylamino-4-thiazolyl)-2-methoxyiminoacetamido]-ceph-3-eme-4-carboxylicacid STEP A: ethyl 2-(2-amino-4-thiazolyl)-2-methoxyiminoacetate

A mixture of 22.5 g of ethyl γ-chloro-α-oximino acetylacetate in 100 mlof methylene chloride was placed in an ice bath and 275 ml of a freshsolution of diazomethane (21.6 g/l) in methylene chloride were slowlyadded with stirring. The mixture was allowed to stand for 5 minutes andthen excess diazomethane was destroyed with a little alumina. Themixture was concentrated to dryness and the residue was chromatographedover silica gel and was eluted with methylene chloride to obtain 11.93 gof ethyl γ-chloro-α-methoxyimino-acetylacetate.

A mixture of 1 g of ethyl γ-chloro-α-methoxyiminoacetylacetate, 3 ml ofabsolute ethanol and 0.42 g of powdered thiourea was stirred at roomtemperature for about 2 hours and was then diluted with 60 ml of etherwhich caused the hydrochloride to crystallize. The mixture was stirredand vacuum filtered and the recovered precipitate was washed and driedto obtain 686 mg of the hydrochloride of ethyl2-(2-amino-4-thiazolyl)-2-methoxyiminoacetate. The latter was dissolvedin 4 ml of water of 50° C. and potassium acetate was added until thesolution has a pH of 6 at which the free amine crystallized. The mixturewas cooled and vacuum filtered and the recovered precipitate was washedwith water and dried to obtain 270 mg of ethyl2-(2-amino-4-thiazolyl)-2-methoxyiminoacetate melting at 161° C. andhaving a syn configuration. RMN spectrum (CDCl₃ 60 MHZ) ppm: 4-(NOCH₃),6.7 (proton of thiazolic ring).

STEP B: ethyl 2-(2-tritylamino-4-thiazolyl)-2-methoxyiminoacetate

2.9 ml of triethylamine were added at -10° C. to a mixture of 4.6 g ofthe product of Step A in 92 ml of methylene chloride and after coolingto -35° C., 6.1 g of tritylchloride were added thereto. The temperaturewas allowed to rise to room temperature in about 21/2 hours and thereaction mixture was washed with water and then with 0.5N hydrochloricacid and finally with aqueous sodium acetate. The mixture was dried andevaporated to dryness. The residue was taken up in ether and thesolution was concentrated to dryness again. The residue was dissolved inmethanol and water and ether were added to cause crystallization. Themixture was vacuum filtered and the recovered precipitate was washedwith ether to obtain 6.15 g of ethyl2-(2-tritylamino-4-thiazolyl)-2-methoxyiminoacetate melting at 120° C.having the syn configuration.

STEP C: 2-(2-tritylamino-4-thiazolyl)-2-methoxyiminoacetic acid

A solution of 7.01 g of the ester of Step B in 35 ml of dioxane washeated to 110° C. on an oil bath and 9 ml of 2N sodium hydroxidesolution was added thereto over 5 minutes. The mixture was stirred atreflux for 30 minutes and the sodium salt crystallized. The mixture wascooled and vacuum filtered and the recovered precipitate was washed withdioxane and then with ether to obtain a first crop of 5.767 g of a salt.The filtrate was concentrated to obtain a second crop of 1.017 g ofsodium salt for a total yield of 6.784 g. 3.06 g of the said salt weredissolved in 65 ml of methylene chloride and 6.5 ml of 2N hydrochloricacid and the solution was washed with water, dried and evaporated todryness to obtain a quantitative yield of2-(2-tritylamino-4-thiazolyl)-2-methoxyiminoacetic acid with a synconfiguration. RMN (DMSO, 60 MHZ) ppm: 3.68 (N--OCH.sub. 3), 6.6 (protonof thiazolic ring).

STEP D:3-acetoxymethyl-7-[2-(2-tritylamino-4-thiazolyl)-2-methoxyiminoacetamido]-ceph-3-eme-4-carboxylicacid

0.78 g of dicyclohexylcarbodiimide was added to a solution of the dryacid of Step C in 30 ml of dry methylene chloride and the mixture wasstirred for an hour at room temperature and was then vacuum filtered toremove dicyclohexyl urea formed. The filtrate was cooled to -10° C. anda solution of 1.01 g of 7-amino-cephalosporanic acid in 13 ml ofmethylene chloride and 0.9 ml of triethylamine was added. Thetemperature returned to room temperature and 1 ml of acetic acid wasadded thereto. The mixture was vacuum filtered and the filtrate waswashed with aqueous hydrochloric acid, dried and concentrated todryness. The residue was taken up in 10 ml of dioxane and 1 ml of waterand 3 ml of a saturated sodium bicarbonate aqueous solution were addedthereto. The mixture was stirred and vacuum filtered and the filtratewas washed and evaporated to dryness. The residue was taken up inmethylene chloride and the solution was washed with 10 ml of water and 5ml of N hydrochloric acid. The mixture was decanted and the organicphase was washed with water, dried and effloresced with ether to obtain1.747 g of raw product. The latter was dissolved in ethyl acetate andwas crystallized by addition of ether to obtain 1.255 g of pure3-acetoxymethyl-7-[2-(2-tritylamino-4-thiazolyl)-2-methoxyiminoacetamido]-ceph-3-eme-4-carboxylicacid in the syn configuration.

EXAMPLE 3A ethyl 2-(2-amino-4-thiazolyl)-2-methoxyiminoacetate STEP A':ethyl 2-acetyl-2-methoxyiminoacetate

234 g of potassium carbonate were added at 10° C. under a nitrogenatmosphere to a mixture of 180 g of raw ethyl2-acetyl-2-hydroxyiminoacetate in 900 ml of pure acetone and then 103 mlof dimethyl sulfate were added thereto. The mixture was stirred at roomtemperature for 3 hours and was then poured into ice and 4 liters ofwater were added. The mixture was extracted with methylene chloride andthe extracts were washed with water, dried and distilled to dryness toobtain 185 g of ethyl 2-acetyl-2-methoxyiminoacetate.

STEP B': ethyl 4-bromo-2-methoxyiminoacetylacetate

200 mg of p-toluene sulfonic acid were added to a mixture of 197 g ofthe product of Step A' in 1 liter of methylene chloride and then 0.1M ofa solution of 191 g of bromine in 200 ml methylene chloride were addedthereto at 20° C. When the reaction had started, the rest of the brominesolution was added over an hour at 20° C. The temperature was raised to25° C. at the end of the reaction and the mixture was washed with icewater and extracted with methylene chloride. The organic extracts weredried and distilled to dryness to obtain 268 g of ethyl4-bromo-2-methoxyiminoacetylacetate.

STEP C': ethyl 2-(2amino-4-thiazolyl)-2-methoxyiminoacetate

A solution of 268 g of the product of Step B' in 270 ml of ethanol wasadded under a nitrogen atmosphere over 30 minutes to a solution of 80 gof thiourea, 270 ml of ethanol and 540 ml of water and the mixture wasstirred for an hour at 20° C. The mixture was cooled to 15° C. and smallamounts of potassium bicarbonate were added to obtain a pH of 5. Themixture was vacuum filtered and the recovered product was washed withwater and dried to obtain 133.8 g of ethyl2-(2-amino-4-thiazolyl)-2-methoxyiminoacetate which was identical to theproduct obtained in Step A of Example 3.

EXAMPLE 43-acetoxymethyl-7-[2-(2-amino-4-thiazolyl)-2-methoxyiminoacetamido]-ceph-3-eme-4-carboxylicacid

A mixture of 0.975 g of the product of Example 3 in 4 ml of 50% aqueousformic acid was stirred at 55° C. for 10 minutes and 4 ml of water wereadded thereto. The mixture was vacuum filtered and the filtrate wasevaporated to dryness under reduced pressure. The residue waseffloresced with 2 ml of ethanol and was vacuum filtered. The productwas washed with ethanol and then with ether to obtain 0.428 g of pure3-acetoxymethyl-7-[2-(2-amino-4-thiazolyl)-2-methoxyiminoacetamido]-ceph-3-eme-4-carboxylicacid with a syn configuration.

RMN (DMSO 60 MHZ) ppm: 2.03 ##STR34## doublet 9.58 J=8 HZ (CONH), 6.76(proton of thiazolic ring).

Analysis: C₁₆ H₁₇ O₇ N₅ S₂ : Calculated: % C, 42.19,% H, 3.76,% N,15.37,% S, 14.08. Found: % C, 42.3,% H, 4.1,% N, 15.2,% S, 13.8.

EXAMPLE 5 diethylamine salt of3-acetoxymethyl-7-[2-(2-tritylamino-4-thiazolyl)-2-methoxyiminoacetamido]-ceph-3-eme-b4-carboxylic acid

The raw acid of Example 3 formed by condensation of 40.8 g of7-amino-cephalosporanic acid and the anhydride ofdicyclohexylcarbodiimide and2-(2-tritylamino-4-thiazolyl)-2-methoxyimino-acetic acid reaction wasdissolved in 350 ml of dioxane and 350 ml of ether and then 33 m l ofdiethylamine were slowly added thereto with stirring. The mixture wasstirred for 20 minutes and the precipitated product was recovered byvacuum filtration, was washed twice with the above ether-dioxane mixtureto obtain 62.6 g of product. The filtrate was concentrated to a syrupyconsistency and was then added to 2.0 liters of ether. The mixture wasstirred and vacuum filtered to obtain 110.3 g of the diethylamine saltof 3-acetoxymethyl7-[2-(2-tritylamino-4-thiazolyl)-2-methoxyiminoacetamido]-ceph-3-eme-4-carboxylicacid in the syn configuration.

EXAMPLE 63-acetoxymethyl-7-[2-(2-amino-4-thiazolyl)-2-methoxyiminoacetamido]-ceph-3-eme-4-carboxylicacid

A mixture of 36 g of the product of Example 5 and 180 ml of 50% aqueousformic acid was stirred at 50° C. for 20 minutes and the triphenylcarbinol formed was removed by vacuum filtration. 180 ml of ethanol wereadded to the filtrate and the mixture was evaporated to dryness underreduced pressure. The residue was taken up in a mixture of 100 ml ofwater and 20 ml of ethanol and was evaporated to dryness again. Theresidue was taken up in 100 ml of water and the mixture was stirred for15 minutes and was vacuum filtered. The recovered product was washedwith water and then ether to obtain 15.6 g of3-acetoxymethyl-7-[2-(2-amino-4-thiazolyl)-2-methoxyiminoacetamido]-ceph-3-eme-4-carboxylicacid which was identical to the product of Example 4.

EXAMPLE 7 sodium3-acetoxymethyl-7-[2-2-amino-4-thiazolyl)-2-methoxyiminoacetamido]-ceph-3-eme-4-carboxylate

A solution of 8 g of sodium bicarbonate in about 20 ml of ethanol wasprogressively added to 45.55 g of pure3-acetoxymethyl-7-[2-(2-amino-4-thiazolyl)-2-methoxyiminoacetamido]-ceph-3-eme-4-carboxylicacid of Example 4 in 100 ml of distilled water and another 80 ml ofethanol and 4.5 g of activated carbon were added thereto. The mixturewas stirred for 5 minutes and was filtered. The filter was rinsed withethanol and the filtrate was evaporated to dryness under reducedpressure. The residue was taken up in 100 ml of ethanol and evaporatedto dryness again. The residue was dissolved in 100 ml of methanol andthe solution was poured into 2 liters of acetone. The mixture wasvigorously stirred and was vacuum filtered. The recovered product wasrinsed with acetone and then ether and dried under reduced pressure toobtain 43.7 g of a white product which rehydrated in air to obtain afinal weight of 45.2 g of sodium3-acetoxymethyl-7-[2-(2-amino-4-thiazolyl)-2-methoxyiminoacetamido]-ceph-3-eme-4-carboxylatewith a specific rotation [α]_(D) ² =+55°±2° (C=0.8% in water). Theproduct had a syn configuration.

RMN (D₂ O-60 MHZ) ppm: 2.01 (COCH₃), doublet at 9.53 J=8 HZ (NHCO), 6.75(proton of thiazolic ring).

Analysis: Calculated: % C, 40,24, % H, 3.38, % N, 14.67, % S, 13.43, %Na, 4.81. Found: % C, 40.3, % H, 3.8, % N, 14.4, % S, 13.3, % N, 4.84.

EXAMPLE 8 3-acetoxymethy-7-[2-(2-tritylamino-4-thiazolyl)-2-(2-propenyl)oxyiminoacetamido]-ceph-3-eme-4-carboxylic acid STEP A: ethyl2-(2-amino-4-thiazolyl)-2-(2-propenyl)oxyiminoacetate

27.5 ml of 2N sodium hydroxide solution was added to an ice-cooledmixture of 9.7 g of ethyl 2-hydroxyimino-4-chloro-acethlacetate, 30 mlof acetone and 9.15 ml of 3-iodopropene and the mixture stood for 1178hours at room temperature. 3.8 g of thiourea were added to the reactionmixture and the mixture was then heated at 60° C. for 15 minutes andstood at room temperature for 45 minutes. The acetone was thenevaporated and methylene chloride, water and potassium carbonate wereadded thereto. The mixture was stirred and decanted. The aqueous phasewas extracted with methylene chloride and the organic extracts weredried and evaporated to dryness to obtain 9.75 of residue. The latterwas chromatographed over silica gel and was eluted with ether to obtain2.7 g of product. The latter was taken up in isopropyl ether and themixture was vacuum filtered. The recovered crystals were rinsed anddried to obtain 783 mg of ethyl2-(2-amino-4-thoazolyl)-2-(2-propenyl)-oxyiminoacetate melting at 100°C. and having the syn configuration.

STEP B: ethyl 2-(2-tritylamino-4-thiazolyl)-2-(2-propenyl)oxyiminoacetate

615 mg of trityl chloride were added at -15° C. to a mixture of 511 mgof the product of Step A, 0.92 ml of dimethylformamide, 1.8 ml ofmethylene chloride and 0.29 ml of triethylamine and the mixture stoodfor 11/2 hours at room temperature. 2 ml of 1N hydrochloric acid andthen 5 ml of water were added thereto and the mixture was decanted. Theorganic solution was dried and concentrated to dryness to obtain 1.28 gof raw ethyl 2-(2-tritylamino-4-thiazolyl-2-(2-propenyl) oxyiminoacetatewith a syn configuration

STEP C: 2-(2-tritylamino-4-thiazolyl)-2-(2-propenyl) oxyimino-aceticacid

A mixture of 1.28 g of the product of Step B, 6.2 ml of dioxane and 3 mlof 2N sodium hydroxide solution was formed at 120° C. and was refluxedfor an hour during which a sodium salt crystallized. The mixture wasvacuum filtered and the recovered product was rinsed with adioxane-ether mixture and dried to obtain 805 mg of the sodium salt. Thelatter was added to 10 ml of methylene chloride and 3 ml of 1Nhydrochloric acid and the mixture was stirred until dissolutionoccurred. The mixture was decanted and the organic phase was dried andconcentrated to dryness. The residue was taken up in ether and themixture was vacuum filtered to obtain 715 mg of2-(2-tritylamino-4-thiazolyl)-2-(2-propenyl) oxyimino-acetic acid with amelting point of 170° C. and a syn configuration.

STEP D:3-acetoxymethyl-7-[2-(2-tritylamino-4-thiazolyl)-2-(2-propenyl)oxyiminoacetamido]-ceph-3-eme-4-carboxylicacid

A mixture of 470 mg of the product of Step C, 5 ml of methylene chlorideand 130 ml of dicyclohexylcarbodiimide was rinsed with a littlemethylene chloride and the mixture was stirred for an hour at roomtemperature. The mixture was vacuum filtered to remove thedicyclohexlurea formed and the filtrate was cooled. A solution of 136 mgof 7-amino-cephalosporanic acid, 2.4 ml of methylene chloride and 0.14ml of triethylamine was added to the filtrate under an inert atmosphereand the mixture sat for 11/2 hours at room temperature 2 ml of 1Nhydrochloric acid and water were added thereto and the mixture wasstirred and decanted. The organic phase was washed with water, dried andevaporated to dryness to obtain 610 mg of raw3-acetoxymethyl-7-[2-(2-tritylamino-4-thiazolyl-2-(2-propenyl)oxyiminoacetamido]-ceph-3-eme-4-carboxylic acid with a synconfiguration.

EXAMPLE 93-acetoxymethyl-7-[2-(2-amino-4-thiazolyl)-2-(2-propenyl)oxyiminoacetamido]-ceph-3-eme-4-carboxylicacid.

A mixture of 610 mg of the product of Example 8 and 3 ml of 50% aqueousformic acid was heated at 60° C. for 15 minutes and 4 ml of water wereadded. The mixture was stirred and vacuum filtered. The filtrate wasrinsed with water and concentrated to dryness under reduced pressure.The residue was taken up in water and was effloresced. The mixture wasvacuum filtered and the product was rinsed to obtain 120 mg of3-acetoxymethyl-7-[2-(2-amino-4-thiazolyl)-2-(2-propenyl)oxyiminoacetamido]-ceph-3-eme-4-carboxylic acid with a melting point ofabout 160° C. and a syn configuration.

    ______________________________________                                        U.V. Spectra (ethanol):                                                       Max. at 236 nm  E.sub.1.sup.1 = 375                                                                         ε = 18,000                              Inflex. towards 252 nm                                                                        E.sub.1.sup.1 = 316                                           Inflex. towards 295 nm                                                                        E.sub.1.sup.1 = 138                                                                         ε = 6,600                               U.V. Spectrum (ethanol -0.1 N HCl):                                           Max at 263 nm   E.sub.1.sup.1 = 380                                                                         ε = 18,300                              Inflex. towards 280 nm                                                                        E.sub.1.sup.1 = 317                                           ______________________________________                                    

RMN (DMSO-90 MHZ) ppm: 2.02 (OAc), 6.68 (proton of thiazolic ring).

EXAMPLE 103-acetoxymethyl-7-[2-(2-tritylamino-4-thiazolyl)-2-ethoxyiminoacetamido]-ceph-3-eme-4-carboxylicacid STEP A: ethyl 2-(2-amino-4-thiazolyl)-2-ethoxyiminoacetate

53 ml of 2N sodium hydroxide solution were added over 30 minutes to amixture of 19.4 g of ethyl γ-chloro-α-oxyiminoacetoacetate in 60 ml ofacetone and 14.3 ml of diethyl sulfate which had set in an ice bath for10 minutes and the mixture was stirred for 40 minutes. 7.6 g of thioureawere added to the reaction mixture which was then heated to 55° C. for20 minutes and the acetone was evaporated. The residue was taken up inethyl acetate and after adding 6.9 g of potassium carbonate thereto, themixture was stirred and decanted. The mixture was extracted with ethylacetate and the extracts were dried and evaporated to dryness. The 17.4g of residue were chromatographed over silica gel and were eluted withether. The product was taken up in isopropyl ether and was vacuumfiltered. The recovered product was rinsed and dried to obtain 2.8 g ofethyl 2-(2-amino-4-thiazolyl)-2-ethoxyiminoacetate melting at 129° C.and having the syn configuration.

STEP B: ethyl 2-(2-tritylamino-4-thiazolyl)-2-ethoxyiminoacetate

3.98 g of trityl chloride were slowly added under an inert atmosphere toa mixture of 3.16 g of the product of Step A, 6 ml of dimethylformamide,12 ml of methylene chloride and 1.89 ml of triethylamine cooled to -15°C. and the mixture stood at 10° C. for 30 minutes and then 3 hours atroom temperature. 13 ml of N hydrochloric acid were added to the mixturewhich was stirred and decanted. The organic phase was washed with Nhydrochloric acid, then with water and was extracted with methylenechloride. The extracts were dried and evaporated to dryness to obtain7.89 g of raw ethyl 2-(2-tritylamino-4 -thiazolyl)-2-ethoxyiminoacetatehaving a syn configuration.

STEP C: 2-(2-tritylamino-4-thiazolyl)-2-ethoxyimino-acetic acid

A mixture of 7.89 g of the product of Step B, 40 ml of dioxane and 19.5ml of 2N sodium hydroxide was heated for one hour at 110° C. and wasvacuum filtered. The recovered product was rinsed with a mixture ofether and dioxane and then with ether and dried to obtain 6.25 g of asodium salt. The latter was taken up in 60 ml of methylene chloride and20 ml of N hydrochloric acid and the two phases were stirred, 20 ml ofmethanol were added thereto and the mixture was decanted. The organicphase was washed with water and was extracted with a methylenechloride-methanol mixture. The extracts were dried and evaporated todryness to obtain 5.85 g of pure2-(2-tritylamino-4-thiazolyl)-2-ethoxyimino-acetic acid with a synconfiguration.

STEP D:3-acetoxymethyl-7-[2-(2-tritylamino-4-thiazolyl)-2-ethoxyiminoacetamido]-ceph-3-eme-4-carboxylicacid.

970 mg of dicyclohexylcarbodiimide were added to a cooled suspension of3.43 g of the acid of Step C in 34 ml of methylene chloride and themixture was rinsed with methylene chloride and was stirred for an hourat room temperature. The mixture was vacuum filtered and the filtratewas cooled to -20° C. and was poured all at once into a solution of 1.02g of 7-amino-cephalosporanic acid in 18 ml of methylene chloride and1.06 ml of triethylamine, cooled to -20° C. The mixture was reheated for11/2 hours and 1.8 ml of acetic acid were added thereto. 9 ml of Nhydrochloric acid were added and the mixture was stirred and decanted.The organic phase was washed with water and was extracted with methylenechloride. The extracts were dried and evaporated to obtain 4.56 g of3-acetoxymethyl-7-[2-(2-tritylamino-4-thiazolyl)-2-ethoxyiminoacetamido]-ceph-3-eme-4-carboxylicacid with a syn configuration.

EXAMPLE 113-acetoxymethyl-7-[2-(2-amino-4-thiazolyl)-2-ethoxyiminoacetamido]-ceph-3-eme-4-carboxylicacid

A mixture of 4.56 g of the product of Example 10 and 23 ml of 50%aqueous formic acid was heated at 55° C. for 15 minutes and was thendiluted with 30 ml of water. The mixture was vacuum filtered and thefiltrate was evaporated to dryness. The residue was taken up in waterand the mixture was stirred and vacuum filtered. The recovered productwas rinsed and dried to obtain 116 mg of impure product. The filtratewas concentrated to obtain a second crop of 674 ml of product for atotal yield of 790 mg.

The 1.063 g of raw product was empasted with 5 ml of water and themixture was heated at 70° C. for 5 minutes and was cooled. The mixturewas stirred for 30 minutes and was vacuum filtered. The product wasrinsed and dried to obtain 8.15 mg of product which was taken up in 2 mlof water and 3 ml of acetone. The mixture was slowly heated and wasvacuum filtered. 3 ml of water were added to the filtrate and themixture was heated to 60° C. and the acetone was removed by bubblingnitrogen therethrough. The mixture was vacuum filtered and the recoveredproduct was rinsed with water, then with ether to obtain 438 mg of3-acetoxymethyl-7-[2-(2-amino-4-thiazolyl)-2-ethoxyiminoacetamido]-ceph-3-eme-4-carboxylicacid with syn configuration.

Analysis: C₁₇ H₁₉ O₇ N₅ S₂ :

Calculated: % C, 43.49, % H, 4.08, % N 14.92, % S, 13.66. Found: % C,44.5, % H, 4.4, % N, 14.8, % S, 13.3.

RMN (60 MHZ DMSO) ppm: 2.05 (OAc), 6.75 (proton or thiazol ring).

EXAMPLE 123-acetoxymethyl-7-[2-(2-tritylamino-4-thiazolyl)-2-(1-methylethoxyimino)acetamido]-ceph-3-eme-4-carboxylicacid STEP A: ethyl 2-ethyl 2-acetyl-2-(1-methyl ethoxyimino)acetate

52 g of potassium carbonate were added to a mixture of 397.8 g of ethyl2-acetyl-2-hydroxyiminoacetate in 200 ml of pure acetone cooled on anice bath and then 25 ml of 2-iodopropane were added thereto over 25minutes. The mixture was stirred for 2 hours and 800 ml of water and 500ml of methylene chloride were added thereto. The mixture was stirred anddecanted and the aqueous phase was extracted with methylene chloride.The mixture was dried and vacuum filtered and the filtrate wasevaporated to obtain 41.5 g of ethyl2-acetyl-2-(1-methylethoxyimino)-acetate.

STEP B: ethyl 4-bromo-2-(1-methylethoxyimino)-acetylacetate

A mixture of 41.5 g of the product of Step A in 190 ml of methylenechloride with traces of p-toluene sulfonic acid was stirred and asolution of 11.9 ml of bromine in 50 ml of methylene chloride was addedthereto over an hour at room temperature. The mixture was stirred andwas then poured into ice water and was decanted. The aqueous phase wasextracted with methylene chloride and the extracts were washed with icewater, dried and evaporated to dryness to obtain 55 g of ethyl4-bromo-2-(1-methylethoxyimino)-acetylacetate.

STEP C: ethyl 2-(2-amino-4-thiazolyl)-2-(1-methylethoxyimino)acetate

A solution of 55 g of the product of Step B in 55 ml of ethanol wasadded over 40 minutes to a solution of 14.9 g of thiourea in 55 ml ofethanol and 105 ml of water and the mixture was stirred at roomtemperature for 21/2 hours. 220 ml of 10% sodium bicarbonate in waterwas added thereto and the mixture was stirred and vacuum filtered. Therecovered product was rinsed and dried to obtain 42.15 g of raw product.The latter was subjected to chromatography with elution with ether. Thefractions rich in the desired product were combined and evaporated todryness. The residue was taken up in isopropyl ether and the mixture wasvacuum filtered. The recovered product was rinsed to obtain 10.75 g ofethyl 2-(2-amino-4-thiazolyl)-2-(1-methylethoxyimino)-acetate in the synconfiguration.

STEP D: ethyl2-(2-tritylamino-4-thiazolyl)-2-(1-methylethoxyimino)-acetate

13.2 g of trityl chloride were slowly added to a cooled mixture of 11 gof the product of Step C, 20 ml of dry dimethylformamide, 40 ml ofmethylene chloride and 6.2 ml of triethylamine and the mixture wasstirred for 21/2 hours, 43 ml of N hydrochloric acid were added theretoand the mixture was stirred and decanted. The organic phase was washedwith 40 ml of water and was extracted with methylene chloride. Theextracts were dried and vacuum filtered. The filtrate was evaporated todryness to obtain 27.7 g of ethyl2-(2-tritylamino-4-thiazolyl)-2-(1-methylethoxyimino)-acetate in the synconfiguration.

STEP E: 2-(2-tritylamino-4-thiazolyl)-2-(1-methylethoxyimino acetic acid

A mixture of 27.7 g of the product of Step D, 150 ml of dioxane and 65ml of 2N sodium hydroxide was refluxed for 2 hours and the mixture wascooled for crystallization of a sodium salt. The mixture was vacuumfiltered and the recovered product was rinsed with a 1-1 ether-dioxanemixture and dried to obtain 16.85 g of raw sodium2-(2-tritylamino-4-thiazolyl)-2-(1-methylethoxyimino)-acetate.

15.9 g of the said salt were dissolved in 15.9 g of dimethylformamide,100 ml of water and 500 ml of methanol and 30 ml of 2N hydrochloric acidwere added thereto. The methanol was evaporated and the mixture wasdiluted with water and was vacuum filtered. The recovered precipitatewas rinsed and dried to obtain 9.8 g of viscous product which was takenup in 220 ml of a 1-1 methylene chloride-methanol mixture. The mixturewas evaporated to dryness and the residue was taken up in ether. Afterefflorescence, the mixture was vacuum filtered and the recovered productwas rinsed with methylene chloride and with water and dried to obtain4.9 mg of 2-(2-tritylamino-4-thiazolyl)-2-(1-methylethoxyimino)aceticacid in the syn configuration and melting at 170° C. For analysis, 300mg of the product were dissolved in 2 ml of methylene chloride and 1 mlof ethanol and the solution was diluted with water and methylenechloride. The mixture was stirred and vacuum filtered and the recoveredcrystals were rinsed with methylene chloride and water dried to obtain230 mg of product for analysis.

Analysis: Calculated: % C, 68.77, % H, 5.34, % N, 8.91, % S 6.8. Found:% C, 68.6, % H, 5.5, % N, 8.8, % S, 6.8.

STEP F:3-acetoxymethyl-7-[2-(2-tritylamino-4-thiazolyl)-2-(1-methylethoxyimino)-acetamido]-ceph-3-eme-4-carboxylicacid.

A solution of 1.62 g of dicyclohexylcarbodiimide in 16 ml of methylenechloride was added under an inert atmosphere to a solution of 4.89 g ofthe product of Step D in 13.5 ml of dimethylformamide in an ice bath andthe mixture was stirred in the ice bath and then vacuum filtered. Therecovered product was rinsed with methylene chloride and dried to remove1.424 g of dicyclohexlurea. The filtrate was cooled in a methanol-icebath and a solution of 1.41 g of 7-aminocephalosporanic acid in 30 ml ofmethylene chloride and 1.45 ml of triethylamine were added thereto. Themixture was stirred for 3 hours at room temperature and after theaddition of 20 ml of N hydrochloric acid, the mixture was stirred anddecanted. The aqueous phase was extracted with methylene chloride andthe extracts were dried and vacuum filtered to obtain 9.05 g of amixture of the starting material and final product. The mixture wastaken up in methylene chloride and was seeded. Crystallization waseffected with stirring and the mixture was vacuum filtered. The crystalswere rinsed and dried to obtain 1.6 g of pure starting material. Thefiltrate was evaporated to dryness and the residue was taken up inisopropyl ether with vigorous stirring to obtain 4.91 g of insolubleviscous 3-acetoxymethyl-7 -[2-(2-tritylamino-4-thiazolyl)-2-(1-methylethoxyimino)-acetamido]-ceph-3-ene-4-carboxylicacid in the syn configuration

EXAMPLE 133-acetoxymethyl-7-[2-(2-amino-4-thiazolyl)-2-(1-methylethoxyimino)-acetamido]-ceph-3-eme-4-carboxylicacid.

A mixture of 4.91 g of raw product of Example 12 in 30 ml of 50% aqueousformic acid was stirred in a water bath at 60° C. and was then dilutedwith water and vacuum filtered. The filter was rinsed with water and theproduct was dried to obtain 1.39 g of triphenylcarbinol. The filtratewas evaporated to dryness and the residue was taken up in water andeffloresced. The mixture was vacuum filtered and the recovered productwas rinsed with water and dried to obtain 800 mg of3-acetoxymethyl-7-[2-(2-amino-4-thiazolyl)-2-(1-methylethoxyimino)-acetamido]-ceph-3-eme-4-carboxylicacid. 972 mg of the product were dissolved in 4 ml of methanol and thesolution was diluted with 20 ml of ether and vacuum filtered. Theproduct was rinsed and dried to obtain 404 mg of the said acid foranalysis in the syn configuration and melting at≃200° C.

Analysis: Calculated: % C, 44.71, % H, 4.38, % N, 14.48, % S, 13.26.Found: % C, 44.5, % H, 4.5, % N, 14.1, % S, 13.2.

RMN spectra (60 MHZ DMSO) ppm: 2.01 (CH₃ CO); doublet at 9.46 J=8 HZ(CONH); 6.7 (proton of thiazole ring).

EXAMPLE 143-acetoxymethyl-7-[2-(2-tritylamino-4-thiazolyl)-2-ethoxyimino)acetamido]-ceph-3-eme-4-carboxylic acid. STEP A: anti isomer of ethyl2-(2-tritylamino-4-thiazolyl)-2-hydroxyiminoacetate

47.6 g of trityl chloride were added in small portions over 30 minutesto a mixture (cooled at -30° C.) of 32.2 g of the anti isomer of ethyl2-(2-amino-4-thiazolyl)-2-hydroxyiminoacetate of Step A of Example 1, 90ml of dry dimethylformamide and 24 ml of triethylamine and the mixturewas left to spontaneous heating for 21/2 hours. Then, 150 ml of 2Nhydrochloric acid and 600 ml of water were added thereto and the mixturewas stirred for 15 minutes, then vacuum filtered. The recoveredprecipitate was empasted 3 times with ether and was then taken up in amixture of methanol, water and triethylamino. The mixture was stirredand then vacuum filtered. The recovered product was rinsed with aqueousmethanol and dried to obtain 60.2 g of the anti isomer of ethyl2-(2-tritylamino-4-thiazolyl)-2-hyroxyiminoacetate, 3.4 g of the saidproduct were recrystallized from a methylene chloride-methanol mixtureto obtain 3 g of pure product melting at 260° C.

STEP B: anti isomer of ethyl2-(2-tritylamino-4-thiazolyl)-2-ethoxyiminoacetate

16.7 ml of ethyl sulfate were added to a mixture of 11.5 g of theproduct of Step A, 5.85 g of potassium carbonate and 25 ml of drydimethylformamide cooled to 15° C. and the mixture stood at roomtemperature for 4 hours. 420 ml of water and 250 ml of ethyl acetatewere added thereto and then was stirred and decanted. The organic phasewas washed with water and extracted with ethyl acetate. The extractswere dried and vacuum filtered and evaporated to dryness. The residuewas taken up in ethanol and crystallization was effected. The solidproduct was rinsed with ethanol, empasted with petroleum ether and driedto obtain 6.6 g of the anti isomer of ethyl2-(2-tritylamino-4-thiazolyl)-2-ethoxyiminoacetate melting at 165° C.797 mg of product were dissolved in a 50-50 methylene chloride-ethanolmixture and the mixture was vacuum filtered. The methylene chloride wasevaporated from the filtrate and the product which crystallized wasrecovered by vacuum filtration was rinsed with ethanol and dried toobtain 596 g of pure product.

STEP C: Anti Isomer of2-(2-tritylamino-4-thiazolyl)-2-ethoxyimino-acetic acid

A mixture of 7.29 g of the product of Step B in 45 ml of dioxane and 9ml of 2N sodium hydroxide was heated on a water bath to 50° C. for 110minutes with stirring and was then crystallization was induced in icedwater and the mixture was vacuum filtered. The recovered product wasrinsed with ether to obtain 4.2 g of the sodium salt which was dissolvedin 50 ml of methylene chloride, 40 ml of water and 11 ml of Nhydrochloric acid. The mixture was stirred and decanted. The mixture wasextracted with methylene chloride and the extracts were washed withwater, dried, vacuum filtered and concentrated to dryness. The residuewas taken up in 50 ml of ether and the solution was stirred and vacuumfiltered. The recovered crystals were rinsed with ether to obtain 3.27 gof the anti isomer of 2-(2-tritylamino-4-thiazolyl)-2-ethoxyimino-aceticacid melting at about 200° C. with decomposition. RMN Spectrum (60 MHZCDCl₃) ppm: 7.66 (proton of thiazole ring), 7.36 (protons of trityl).

STEP D: anti isomer of3-acetoxymethyl-7-[2-(2-tritylamino-4-thiazolyl)-2-ethoxyimino)acetamido]-ceph-3-eme-4-carboxylicacid.

1.17 ml of isobutyl chloroformate was added dropwise under an inertatmosphere to a mixture of 4.1 g of the product of Step C, 36 ml oftetrahydrofuran, 27 ml of methylene chloride and 0.99 ml ofN-methyl-morpholine cooled to -20° C. and after standing at -20° C. for3 minutes, the mixture was cooled to -35° C. and a solution of 2.45 g of7-aminocephalosporanic acid in 45 ml of methylene chloride and 252 ml oftriethylamine were added thereto. The mixture stood for 21/2 hours withspontaneous heating and was then evaporated to dryness. The residue wastaken up in a mixture of methylene chloride, water and N hydrochloricacid with a pH of 1-2 and the mixture was extracted with methylenechloride. The organic extracts were washed with water, dried and vacuumfiltered. The filtrate was evaporated to dryness and the residue wastaken up in ethyl acetate. The solution was diluted with isopropyl etherand was stirred and vacuum filtered. The recovered product was rinsedwith isopropyl ether and dried to obtain 4.87 g of the anti isomer of3-acetoxymethyl-7-/2-(2-tritylamino-4-thiazolyl)-2-ethoxyiminoacetamido/-ceph-3-eme-4-carboxylicacid. The said product was dissolved in 10 ml of not ethyl acetate andthe solution was slowly diluted with isopropyl ether. The mixture wasstirred and was vacuum filtered and the recovered product was rinsedwith isopropyl ether and dried to obtain 4.53 g of purified product.

EXAMPLE 15 anti isomer of3-acetoxymethyl-7-[2-(2-amino-4-thiazolyl)-2-ethoxyiminoacetamido]-ceph-3-eme-4-carboxylicacid.

A mixture of 4.27 g of the purified product of Example 14 in 20 ml of50% aqueous formic acid was stirred. on a water bath heated at 60° C.for 20 minutes and was then cooled and diluted with water. The mixturewas stirred for 10 minutes and was vacuum filtered to removetriphenylcarbinol which was rinsed with water to obtain a final weightof 1.44 g. The filtrate was added to ethanol and was evaporated todryness. The residue was dissolved in ethanol and the solution was againevaporated to dryness under reduced pressure. The residue was added to30 ml of water and the mixture was stirred for an hour in an ice bathand was then vacuum filtered. The recovered product was rinsed and driedto obtain 2.06 g of3-acetoxymethyl-7-[2-(2-amino-4-thiazolyl)-2-ethoxyiminoacetamido]-ceph-3-eme-4-carboxylicacid. The product was purified by dissolving it in 5 ml of water and 5ml of 10% aqueous sodium bicarbonate solution and the mixture was vacuumfiltered. The filter was rinsed with water and the filtrate was adjustedto a pH of 3-4 by dropwise addition of formic acid. After standing atroom temperature for 12 hours, the mixture was vacuum filtered and therecovered crystals were rinsed with water and dried to obtain 1.73 g ofpure product melting at about 200° C. with decomposition.

RMN Spectrum (60 MHZ DMSO) ppm: 2.04 (CH₃ CO--) and 7.5 (proton ofthiazole ring).

EXAMPLE 16 anti isomer of3-acetoxymethyl)-7-[2-(2-tritylamino-4-thiazolyl-2-(1-methylethoxyimino)-acetamido]-ceph-3-eme-4-carboxylicacid. STEP A: anti isomer of ethyl2-(2-tritylamino-4-thiazolyl)-2-(1-methylethoxyimino)-acetate

A mixture of 6.86 g of the anti isomer of ethyl2-(2-tritylamino-4-thiazolyl)-2-hydroxyiminoacetate of Example 14, 3.51g of potassium carbonate in 15 ml of dimethylformamide and 7.7 ml ofisopropyl iodide was held under an argon atmosphere and stirring for41/2 hours and then 250 ml of distilled water and 150 ml of ethylacetate were added thereto with stirring. The mixture was decanted andthe aqueous phase was washed with water and was extracted with ethylacetate. The organic phase was dried, vacuum filtered and evaporated todryness. The residue was taken up in ethanol and crystallization startedafter seeding. The mixture was vacuum filtered and the precipitate wasrinsed with ethanol and empasted with petroleum ether to obtain 3.26 gof the anti isomer of ethyl2-(2-tritylamino-4-thiazolyl)-2-(1-methylethoxyimino)-acetate melting at182° C.

STEP B: anti isomer of2-(2-tritylamino-4-thiazolyl)-2-(1-methylethoxyimino)-acetic acid

A mixture of 6.8 g of the product of Step A, 41 ml of dioxane and 8.15ml of 2N sodium hydroxide was heated on a water bath at 55° C. for 2hours and was then cooled. 9.5 ml of 2N hydrochloric acid were added toobtain a pH of 2-3 and the dioxane was distilled. Crystallization waseffected and water diluted the mixture under stirring. The recoveredprecipitate was vacuum filtered, rinsed with water, empasted with etherand dried to obtained 5.87 g of the anti isomer of2-(2-triethylamino-4-thiazolyl)-2-(1-methylethoxyimino)-acetic acidmelting at 240° C. (decomp.).

RMN (CDCl₃, 60 MHZ) ppm: =7.66 (proton of thiazolic ring) and 7.31(trityl group).

STEP C: anti isomer of3-acetoxymethyl-7-[2-(2-tritylamino-4-thiazolyl)-2-(1-methylethoxyimino)-acetamido]-ceph-3-eme-4-carboxylicacid

A mixture of 5.66 g of the product of Step B, 48 ml of tetrahydrofuran,48 ml of methylene chloride and 1.32 ml of N-methyl-morpholine washeated until dissolution and was then cooled to -20° C. 1.56 ml ofisobutyl chloroformate were added and the mixture stood at -10° to -20°C. for 10 minutes and was then cooled to -35° C. A mixture of 3.26 g of7-amino-cepholsporanic acid, 60 ml of methylene chloride and 3.36 ml ofdry triethylamine were added all at once and the mixture was heated withstirring for 31/2 hours. The solvents were distilled and the residue wastotally dissolved in the ethyl acetate and the solution was diluted withisopropyl ether and was stirred and vacuum filtered. The recoveredprecipitate was rinsed with isopropyl ether and dried to obtain 5.42 gof product, 5.82 g of the latter was dissolved in 20 ml of hot ethylacetate and the solution was diluted with 200 ml of isopropyl ether,then vacuum filtered. The precipitate was dried to obtain 4.82 g of theanti isomer of3-acetoxymethyl-7-[2-(2-tritylamino-4-thiazolyl)-2-(1-methylethoxyimino)acetamido]-ceph-3-eme-4-carboxylicacid.

EXAMPLE 17 anti isomer of3-acetoxymethyl-7-[2-(2-amino-4-thiazolyl)-2-(1-methylethoxyimino)-acetamido]-ceph-3-eme-4-carboxylic acid

A mixture of 3.62 g of the product of Example 16 and 16 ml of 50%aqueous formic acid was held at 60° C. for 20 minutes and was thencooled at 20° C. 16 ml of water were added and the mixture was stirredand vacuum filtered. The precipitate was rinsed and dried to obtain 1.23g of triphenylcarbinol. The filtrate was evaporated to dryness underreduced pressure and the residue was taken up in ethanol which solutionwas evaporated to dryness under reduced pressure. The residue was takenup in water and the mixture was stirred and then cooled to inducecrystallization. The mixture was vacuum filtered and the gummy crystalswere rinsed with water and dried to obtain 1.68 g of raw product.

A solution of 2 g of the latter product in 5 ml of water and 5 ml of 5%aqueous sodium bicarbonate solution was stirred for 10 minutes and themixture was vacuum filtered. The precipitate was rinsed with water andthe pH of the filtrate was adjusted to 3 with formic acid. The mixturewas stirred for an hour in an ice bath and the mixture was vacuumfiltered,. the recovered gum was rinsed with water and was dissolved in10 ml of hot ethanol. The solution was cooled with ice water and wasvacuum filtered. The precipitate was rinsed with ethanol and then withether to obtain 748 of the anti isomer of3-acetoxymethyl-7-[2-(2-amino-4-thiazolyl)-2-(1-methylethoxyimino)-acetamido]-ceph-3-eme-4-carboxylicacid. A formation of a crystalline deposit formed in the filtrate whichwas diluted with ether without precipitation. The mixture was vacuumfiltered and the recovered crystals were rinsed with a mixture of etherand ethanol and then with ether to obtain another 177 mg of pure productwhich melted at about 200° C. with decomposition.

RMN (DMSO 60 HMZ) ppm:=7.46 (proton of thiazole ring) and 4.43 (tertiaryproton of isopropyl).

EXAMPLE 18 anti isomer of3-acetoxymethyl-7-[2-(2-tritylamino-4-thiazolyl)-2-(2-propenyloxyimino)-acetamido]-ceph-3-eme-4-carboxylicacid STEP A: anti isomer of ethyl 2-(2-tritylamino-4-thiazolyl)2-(2-propenyloxyimino)-acetate

A mixture of 6.86 g of the anti isomer of ethyl2-(2-tritylamino-4-thiazolyl)-2-hydroxyiminoacetate [Step A of Example14], 3.51 g of potassium carbonate, 15 ml of dimethylformamide and 7 mlof allyl iodide was stirred under an inert atmosphere at roomtemperature for 5 hours and then 250 ml of water and 150 ml of ethylacetate were added thereto with stirring. The mixture was decanted andthe aqueous phase was washed and reextracted with ethyl acetate. Theextracts were dried, vacuum filtered and evaporated to dryness. Theresidue was taken up in ethanol and crystallization was started. Themixture was cooled in an ice bath with stirring for half an hour and wasthen vacuum filtered. The crystals were rinsed with ethanol to obtain4.72 g of the anti isomer of ethyl2-(2-tritylamino-4-thiazolyl)-2-(2-propenyloxyimino)-acetate. 215 mg ofthe product were dissolved in 2 ml of ethanol and 2 ml of methylenechloride and the mixture was filtered. The filtrate was concentrated anddiluted with ethanol. Crystallization was effected in an ice bath andthe mixture was vacuum filtered. The crystals were rinsed with ethanoland dried to obtain 70 mg of the pure product which melted at 90° C.(pasty) and 160° C. (pure).

STEP B: anti isomer of2-(2-tritylamino-4-thiazolyl)-2-(2-propenyloxyimino)-acetic acid

A mixture of 3.71 g of the product of Step A, 22 ml of dioxane and 4.5ml of 2N sodium hydroxide was heated in a water bath at 55° C. for 110minutes and was then cooled. 5.25 ml of 2N hydrochloric acid were addedthereto to adjust the pH to 2 and the dioxane was evaporated. The gummyresidue was diluted with water and the mixture was cooled in an ice bathand was vacuum filtered. The recovered precipitate was rinsed with waterand empasted 3 times with ether to obtain 2.85 g of the anti isomer of2-(2-tritylamino-4-thiazolyl)-2-(2-propenyloxyimino)-acetic acid meltingat 198° C. (decomp.) RMN (CDCl₃ 90 MHZ) ppm:=7.64 (proton of thiazolering) and 7.27 (trityl proton).

STEP C: anti isomer of3-acetoxymethyl-7-[2-(2-tritylamino-4-thiazolyl)-2-(2-propenyloxyimino)-acetamido]-acetamido]-ceph-3-eme-4-carboxylicacid

0.78 ml of isobutyl chloroformate were added at -20° C. to a mixture of2.82 g of the product of Step B, 24 ml of dry tetrahydrofuran and 24 mlof methylene chloride to which 0.66 ml of N-methyl-morpholine had beenadded and the mixture was stirred at -20° C. for 3 minutes. The mixturewas then cooled to -35° C. and a solution of 1.63 g of7-aminocephalosporanic acid, 30 ml of methylene chloride and 1.68 ml oftriethylamine was added thereto. The mixture was returned too roomtemperature during 3 hours and was evaporated to dryness. The residuewas taken up in methylene chloride, and 50 ml of water and 15 ml of Nhydrochloric acid were added. The mixture was stirred, decanted, andwashed with water. The mixture was extracted with methylene chloride andthe extracts were dried and vacuum filtered. The filter was rinsed withmethylene chloride and the filtrate was evaporated to dryness. Theresidue was taken up in ethyl acetate and the solution was diluted withisopropyl ether, was stirred and was vacuum filtered. The recoveredprecipitate was rinsed with isopropyl ether to obtain 3.08 g of the antiisomer of 3-acetoxymethyl-7-[2-(2-tritylamino-4-thiazolyl)-2-(2-propenyloxyimino)-acetamido]-ceph-3-eme-4-carboxylic acid. 3.59 g of the said product were dissolved in 15 ml ofethyl acetate and the solution was diluted with isopropyl ether, wasstirred and was vacuum filtered to obtain 3.33 g of the said purifiedproduct.

EXAMPLE 19 anti isomer of3-acetoxymethyl-7-[2-(2-amino-4-thiazolyl)-2-(2-propenyloxyimino)-acetamido]-ceph-3-eme-4-carboxylicacid

A mixture of 2.53 g of the product of Example 18 in 11.5 ml of a 50%aqueous formic acid solution was heated at 60° C. for 20 minutes and wasthen diluted with water and cooled to room temperature. The mixture wasvacuum filtered and the solids were rinsed with water and dried toobtain 963 mg of triphenylcarbinol. The filtrate and wash waters wereevaporated to dryness under reduced pressure and the residue was takenup in ethanol. The solution was evaporated to dryness and the gumresidue was taken up in 15 ml of water. The mixture effloresced and wasvacuum filtered. The solid was rinsed with water and dried to obtain1.275 g of product. A suspension of 1.63 g of the said raw product in 15ml of ethanol was refluxed and was then vacuum filtered. The precipitatewas rinsed with ethanol to obtain 877 mg of insoluble product. 20 ml ofether were added to the filtrate and the mixture was vacuum filtered.The solids were rinsed with a 1-1 ethanol-ether mixture and were driedto obtain a second crop of 97 mg. The filtrate was concentrated at whichtime crystallization occurred. The product was recovered by vacuumfiltration, was rinsed with ethanol then with ether and dried to obtain162 mg of pure anti isomer of 3-acetoxymethyl-7-[2-(2-amino-4-thiazolyl)-2-(2-propenyloxyimino-acetamido]-ceph-3-eme-4-carboxylicacid with a melting point of 180° C. (pasty).

RMN (DMSO 60 MHZ) p.p.m.=7.48 (thiazole ring proton) and 2.04 (proton ofacetyl).

EXAMPLE 20 syn isomer of3-acetoxymethyl-7-[2-(2-amino-4-thiazolyl)-2-methoxyiminoacetamido]-ceph-3-eme-4-carboxylicacid STEP A: syn isomer of ethyl2-(2-chloroacetamido-4-thiazolyl)-2-methoxyiminoacetate

A mixture of 45.8 g of the syn isomer of ethyl2-(2-amino-4-thiazolyl)-2-methoxyiminoacetate (Step A of Example 3) in200 ml of methylene chloride was heated to distill 20 ml and was thencooled to 10° C. 50 ml of pyridine were added to the mixture followed bythe addition of 41 g of monochloroacetic acid anhydride and the mixturewas heated until dissolution occurred. The mixture stood at 20° C. undernitrogen for 6 hours and 5 ml of water were added with stirring. Themixture was poured into 300 ml of iced 2N hydrochloric acid and was thendecanted. The aqueous phase was extracted with methylene chloride andthe organic extracts were washed with water, with sodium bicarbonatesolution and then with water, was dried, treated with carbon black andwas concentrated. 300 ml of isopropyl ether were added thereto and theproduct crystallized. The mixture was concentrated to obtain a thickpaste which was iced and vacuum filtered. The solid was washed withisopropyl ether and dried to obtain 45.4 g of the syn isomer of ethyl2-(2-chloroacetamido-4-thiazolyl)-2-methoxyiminoacetate melting at 113°C. A sample was crystallized from an isopropyl ether-methylene chloridemixture to obtain a product melting at 118° C.

RMN (CDCl₃ 60 MHz) (a) triplet center about 1.38 ppm J=7 Hz; (b)singulet 4.05 ppm; (c) quadruplet center about 4.44 ppm J=7 Hz; (d)singulet 4.33 ppm; (e) singulet 7.27 ppm; and (f) singulet 9.95 ppm.##STR35##

STEP B: syn isomer of2-(2-chloroacetamido-4-thiazolyl)-2-methoxyimino-acetic acid

30 ml of pure sodium carbonate solution were added at 20° C. under anitrogen atmosphere to a mixture of 46 g of the product of Step A in 230ml of absolute ethanol and the product dissolved. Crystallization of asodium salt commenced and then the medium became a mass. After 16 hours,the mixture was vacuum filtered and the solid was washed with ethanol.The solid was dissolved in water and after icing, 100 ml of 2Nhydrochloric acid were added thereto. The mixture was saturated withsodium chloride and was then extracted with ethyl acetate containing 10% ethanol. The extracts were dried, treated with carbon black anddistilled to dryness under reduced pressure. The water was extrainedwith benzene. The residue was taken up in methylene chloride and thesolution was evaporated to dryness. Again, the residue was taken up inmethylene chloride and the solution was iced and vacuum filtered. Thesolid was washed with methylene chloride and dried to obtain 34.5 g ofthe syn isomer of 2-(2-chloroacetamido-4-thiazolyl)-2-methoxyimino-acetic acid melting at about 200° C. The product waspurified by crystallization from an acetone-isopropyl ether mixture.

Analysis: C₈ H₈ O₄ N₃ ClS; molecular weight=277.68; Calculated: %C,34.60, %H, 2.90, %N, 15.13, %Cl, 12.77, %S, 11.55. Found: %C, 34.8, %H,2.8, %N, 14.8, %Cl, 12.6, %S, 11.5.

RMN (DMSO 60 MHz): (a) singulet 3.92 ppm; (b) singulet 4.38 ppm; (c)singulet about 5 ppm; (d) singulet 7.58 ppm; and (e) singulet 12.6 ppm.##STR36##

STEP C: syn isomer of3-acetoxymethyl-7-[2-(2-chloroacetamido-4-thiazolyl)-2-methoxyimino)-acetamido]-ceph-3-eme-4-carboxylicacid

8 ml of triethylamine were added at 5° C. to a mixture of 15.3 g of theproduct of Step B in 80 ml of methylene chloride and then 3.8 ml ofthionyl chloride and 26 ml of methylene chloride were added thereto at0° C. The mixture stood for 15 minutes at 0° C. and then 7 ml oftriethylamine were added. A mixture of 13.6 g of 7-amino-cephalosporanicacid in 100 ml of methylene chloride and 14 ml of triethylamine wereadded thereto at 0° C. under nitrogen and after returning thetemperature to 20° C., the mixture was stirred for an hour. The mixturewas evaporated to dryness under reduced pressure at 30°-35° C. and theresidue was dissolved in 250 ml of water. The solution was treated withcarbon black and 50 ml of 2N hydrochloric acid were added. Theprecipitate was recovered by vacuum filtration and was washed with waterand was then suspended in 80 ml of ethanol. 7 ml of triethyl amine wereadded at 5° C. and then 15 ml of 4N sulfuric acid were added all at oncewith stirring at 5° C. Crystallization was effected for 15 minutes andthe mixture was vacuum filtered. The product was washed with ethanol byempasting and then with ether and dried under reduced pressure to obtain18.6 g of the syn isomer of3-acetoxymethyl-7-[2-(2-chloroacetamido-4-thiazolyl)-2-methoxyiminoacetamido]-ceph-3-eme-4-carboxylicacid with a specific rotation of [α]_(D) ²⁰ =+26°±1° (C=1% indimethylformamide).

RMN (DMSO 60 MHz): (a) singulet 2.03 ppm; (b) singulet 3.90 ppm; (c)singulet 4.38 ppm; and (d) singulet 7.45 ppm. ##STR37##

STEP D: syn isomer of3-acetoxymethyl-7-[2-(2-amino-4-thiazolyl)-2-methoxyiminoacetamido]-ceph-3-eme-4-carboxylicacid

1 g of potassium bicarbonate was added at 20° C. to a suspension of 5.32g of the product of Step C in 10.6 ml of water and 912 mg of thioureaand after dissolution, the mixture was stirred at 20° C. under nitrogenfor 6 hours. The gummy precipitation started after about 90 minutes andthen 30 ml of water and 3 ml of formic acid were added thereto. Themixture was cooled to 5° C. and was vacuum filtered. The product waswashed with water containing 10% formic acid and was dissolved at 5° C.in 30 ml of water containing triethylamine. 3 ml of formic acid wereadded at 5° C. and the mixture was vacuum filtered. The precipitate wasempasted with water containing formic acid and the dark brown gum waseliminated. The combined aqueous phases were treated with carbon blackto obtain a clear yellow solution which was saturated with ammoniumsulfate and then vacuum filtered. The precipitate was empasted withwater, was vacuum filtered and washed with water to obtain a first crop.The mother liquors were saturated with ammonium sulfate to effectprecipitation and were vacuum filtered then washed with water to obtaina second crop. The two crops were combined and were taken up in ethanol.The mixture was stirred at 20° C. for an hour and stood for 16 hours at0° C. The mixture was vacuum filtered and the product was washed withethanol and then with ether and was dried under reduced pressure toobtain 3.47 g of the syn isomer of3-acetoxymethyl-7-[2-(2-amino-4-thiazolyl)-2-methoxyiminoacetamido]-ceph-3-eme-4-carboxylic acid identical to that of Examples 4 and 6.

RMN (DMSO 60 MHz): (a) singulet 2.03 ppm; (b) singulet 3.55 ppm; (c)doublet 5.19 ppm J=5 Hz; and (d) singulet 6.8 ppm. ##STR38##

EXAMPLE 21 diethylamine salt of syn isomer of3-acetoxymethyl-7-[2-(2-tritylamino-4-thiazolyl)-2-methoxyiminoacetamido]-ceph-3-eme-4-carboxylicacid STEP A: ethyl 2-acetyl-2-methoxyiminoacetate

6.1 kg of potassium carbonate were added at 20°-25° C. to a mixture of4.69 kg of ethyl 2-acetyl-2-hydroxyiminoacetate (equal to 4.21 kg ofpure product) in 21 liters of anhydrous pure acetone and after stirringthe suspension for 10 minutes, 3.72 kg of dimethyl sulfate were added at20°-25° C. The mixture was poured into 126 liters of demineralized waterand was extracted 4 times with 5 liters and then with 2 liters ofmethylene chloride. The combined extracts were washed with 10 liters ofdemineralized water, were dried and vacuum filtered. The filter wasrinsed with 2 liters of methylene chloride and the filtrate wasevaporated to dryness under reduced pressure to obtain 4.88 kg of ethyl2-acetyl-2-methoxyiminoacetate with an RF=0.7 (thin layerchromatography-9-1 methylene chloride-ethyl acetate eluant). The productwas identical to that product in Step A of Example 3.

STEP B: ethyl 4-bromo-2-methoxyimino-acetylacetate

A solution of 2.96 kg of bromine in 3.5 liters of methylene chloride wasadded over 30 minutes at 22° C.±1° to a solution of 3.53 kg of theproduct of Step A in 18.6 liters of methylene chloride and 3.5 g ofp-toluene sulfonic acid and hydrobromic acid was released for 15minutes. The mixture was stirred for 45 minutes at 22° C. The organicphase was washed twice with 14 liters of demineralized water and thewash water was extracted twice with 3.5 liters of methylene chloride.The combined organic phase was dried, filtered, rinsed with methylenechloride and evaporated to dryness under reduced pressure to obtain 4.73kg of ethyl 4-bromo-2-methoxyimino-acetylacetate.

STEP C: syn isomer of ethyl2-(2-amino-4-thiazolyl)-2-methoxyiminoacetate

A mixture of 1.43 kg of thiourea, 3.55 liters of ethanol and 7.1 litersof demineralized water was stirred for 10 minutes at 20° C. and then amixture of 4.730 kg of the product in Step B in 3.55 liters of ethanolwas added thereto at 20°-25° C. The mixture was stirred for 3 hours at20°-25° C. and was then cooled to 15°-25° C. The mixture was neutralizedto a pH of 7 with about 1.6 liters of 22°Be ammonium hydroxide solutionand the mixture was stirred for 15 minutes at 20°-25° C. and was vacuumfiltered. The recovered precipitate was washed 5 times with 1.8 litersof demineralized water, and dried to obtain 2.947 kg of the syn isomerof ethyl 2-(2-amino-4-thiazolyl)-2-methoxyiminoacetate melting at 162°C. The product was identical to that of Step A of Example 3.

STEP D: syn isomer of ethyl2-(2-tritylamino-4-thiazolyl)-2-methoxyiminoacetate

A mixture of 3.41 kg of the product of Step C, 17 liters of methylenechloride and 2.275 liters of triethylamine was stirred for 15 minutesand then 4.55 kg of tritylchloride were added at 20°-25° C. over onehour with stirring under nitrogen. The mixture was stirred for 20 hoursat 20°-25° C. under nitrogen during which triethylamine hydrochloridecrystallized in the mixture was washed with 10.2 liters of iced 0.5Nhydrochloric acid and twice with 10.2 liters of iced demineralized waterand the was waters were extracted with 1.7 liters of methylene chloride.The combined organic extracts were dried and filtered and the filter waswashed with 1.7 liters of methylene chloride. The filtrates wereevaporated to dryness under reduced pressure below 50° C. to obtain8.425 kg of raw product. The latter was dissolved at 20°-25° C. in 8.4liters of methanol and the solution was stirred for an hour at 20°-25°C. after the addition of 2.8 liters of demineralized water to inducecrystallization. The mixture was stirred for another hour and was vacuumfiltered. The product was empasted twice with 1.7 liters of methanolcontaining 25% water and was dried at 40° C. to obtain 7.165 kg of thesyn isomer of ethyl 2-(2-tritylamino-4-thiazolyl)-2-methoxyiminoacetatewhich was identical to the product of Example 3, Step B.

STEP E: syn isomer of sodium salt of2-(2-tritylamino-4-thiazolyl)-2-methoxyiminoacetamido

A mixture of 4.175 kg of the product of Step D in 20.9 liters of ethanolwas refluxed under nitrogen with stirring to obtain totaldissolution at55° C. and then 5.235 liters of about 2N sodium hydroxide were addedthereto at reflux under nitrogen. Crystallization was rapid and themixture was stirred for one hour at reflux under nitrogen and was cooledto 20°-25° C. and held there for 2 hours. The mixture was vacuumfiltered and the solid product was washed 4 times with 2.1 liters ofethanol and was dried to obtain 4.02 kg of the syn isomer of sodium saltof 2-(2-tritylamino-4-thiazolyl)-2-methoxyiminoacetamido.

STEP F: syn isomer of2-(2-tritylamino-4-thiazolyl)-2-methoxyimino-acetic acid

2 liters of about 1N hydrochloric acid were added in 2 minutes at20°-25° C. under nitrogen to a mixture of 500 g of the product of Step E(440 g of dry product) in 2.5 liters of methylene chloride and themixture was stirred under a nitrogen atmosphere for 2 hours. The mixturewas decanted and the organic phase was washed 3 times with 2 liters ofdemineralized water and the wash waters were extracted with 1 liter ofmethylene chloride. The combined organic phases were dried, treated with25 g of carbon black, vacuum filtered, rinsed with methylene chlorideand evaporated to dryness to obtain 481 g of raw product. The latter wastaken up in 2.1 liters of isopropyl ether and the solution was vacuumfiltered. The solid product was washed twice with 420 ml of isopropylether and was dried under reduced pressure to obtain 424.6 g of the synisomer of 2-(2-tritylamino-4-thiazolyl)-2-methoxyiminoacetic acididentical to the product of Step C of Example 3.

STEP G: syn isomer of diethylamine3-acetoxymethyl-7-[2-(2-tritylamino-4-thiazolyl)-2-methoxyiminoacetamido]-ceph-3-eme-4-carboxylate

1200 ml of methylene chloride were added to 200 g of the product of StepF and the mixture was refluxed with stirring under an inert atmosphere.Then, 600 ml of methylene chloride were distilled at normal pressure andafter cooling to 18°-20° C., a solution of 54 g ofdicyclohexylcarbodiimide in 54 ml of methylene chloride were added at18°-20° C. The mixture was stirred under an inert atmosphere for an hourat 18°-20° C. and then an extemporaneously prepared solution of 61.4 gof 7-amino-cephalosporanic acid in 900 ml of methylene chloride and 63ml of triethylamine were added at that temperature over 15 minutes. Themixture with a pH of 6.5 to 7 was stirred at 20° C. for 90 minutes and50 ml of acetic acid were added. The mixture was stirred another 15minutes at 20° C. and was then vacuum filtered. The filter was rinsed 4times with 200 ml of methylene chloride and the filtrate was washed 3times with 400 ml of demineralized water and was then dried overmagnesium sulfate and was vacuum filtered. The filter was washed with200 ml of methylene chloride and the filtrate was evaporated to drynessunder an inert atmosphere with reduced pressure. The dry oil residue wasdissolved in 700 ml of dioxane with stirring under an inert atmosphereat 20°-25° C. and the solution was evaporated to dryness under an inertatmosphere with reduced pressure at a temperature below 30° C. Theresidue was taken up in 300 ml of a dioxane-methylene chloride mixtureand then 500 ml of ether were added at 20°±2° C. followed by theaddition of 52 ml of diethylamine. After about 10 minutes,crystallization occured and the mixture stood at 20° C. under an inertatmosphere for an hour. The mixture was vacuum filtered and the solidproduct was rinsed 3 times with 100 ml of a dioxane-ether solution andwas dried to obtain 113.6 g of the syn isomer of diethylamine3-acetoxymethyl-7-[2-(2-tritylamino-4-thiazolyl)-2-methoxyimonoacetamido]-ceph-3-eme-4-carboxylate. 3.25 liters of isopropyl ether were added over 30minutes to the filtrate and the mixture was stirred for 15 minutes andwas then vacuum filtered. The solid product was rinsed twice with 400 mlof isopropyl ether and was dried under reduced pressure to obtain 182 gof product identical to that of Example 5.

EXAMPLE 22 syn isomer of3-acetoxymethyl-7-[2-(2-amino-4-thiazolyl)-2-methoxyiminoacetamido]-ceph-3-eme-4-carboxylicacid

A mixture of 182 g of the product of Example 21 in 347 ml of formic acidand 87 ml of demineralized water was stirred under an inert atmosphereat 28°-30° C. until there was total dissolution followed bycrystallization of triphenyl carbinol and the mixture was stirred foranother 21/2 hours at 28°-30° C. under argon. Precipitation was effectedwith 1740 ml of demineralized water and 847 g of ammonium sulfate for 15minutes with stirring and then the mixture was stirred for 30 minutesand was then vacuum filtered. The solid product was rinsed twice with174 ml of demineralized and was dried under reduced pressure at 25°-30°C. to obtain 147 g of a mixture of product and triphenyl carbinol. Theraw product was empasted with ether for an hour at 18°-20° C. and wasthen vacuum filtered. The product was rinsed twice with 147 ml of etherand then dried at 25°-30° C. to obtain 89 g of product which wasempasted with stirring under nitrogen with 445 ml of ethanol. Thesuspension was stirred for an hour at 45°-50° C. and one hour at 18°-20°C. and was then vacuum filtered. The product was rinsed twice with 45 mlof ethanol and was dried under reduced pressure at 20° C. to obtain76.85 g of the syn isomer of3-acetoxymethyl-7-[2-(2-amino-4-thiazolyl)-2-methoxyiminoacetamido]-ceph-3-eme-4-carboxylicacid.

The latter product in 230 ml of acetic acid was stirred for 15 minutesunder nitrogen and then 77 ml of demineralized water were added theretofollowed by the addition of about 700 ml of water. The mixture wasstirred for an hour at 18°-20° C. and 269 g of ammonium sulfate wereadded over 10 minutes. The mixture stood for 15 minutes and 3.85 ofcarbon black were added. The mixture was stirred for 15 minutes and wasthen vacuum filtered. The product was rinsed with 77 ml of demineralizedwater containing 25% of acetic acid and 154 ml of formic acid were addedwith stirring at 18°-20° C. to the filtrate. A seed of the final productwas added to induce crystallization by scratching and the mixture wasstirred for 2 hours at 18°-20° C. and then 2 hours at 0° to 5° C. Themixture was vacuum filtered and the product was washed 4 times with 77ml of demineralized water containing 5% formic acid and was dried underreduced pressure at 20°-25° C. to obtain 49.45 g of the said product inthe form of its formate. The latter was empasted with 250 ml of ethanolwith stirring for an hour at 45°-50° C. and then 1 hour at 18°-20° C.The mixture was vacuum filtered and the product was rinsed twice with 50ml of ethanol and dried at 20° C. under reduced pressure and then at35°-40° C. for 10 to 15 hours to obtain 45.45 g of the said product witha specific rotation [α]_(D) ²⁰ =+64.5° (C=0.5% in water with 0.5%NaHCO₃). The product was identical to that of Examples 4, 6 and 20.

EXAMPLE 23 cristallize syn isomer of sodium3-acetoxymethyl-7-[2-(2-amino-4-thiazolyl)-2-methoxyiminoacetamido]-ceph-3-eme-4-carboxylate

A solution of 19.8 g of the product of Example 22 in 65 ml of a molarsolution of sodium acetate in methanol stood at room temperature for 35minutes for crystallization and then 40 ml of ethanol were added theretoover an hour. The mixture was stirred in an ice bath for 21/2 hours andwas then vacuum filtered. The product was washed twice with 10 ml of a1-1 methanol-ethanol mixture and twice with 10 ml of ethanol and finallytwice with 20 ml of ether. The product was dried for 2 hours underreduced pressure at 45° C. and 48 hours under reduced pressure in thepresence of sulfuric acid to obtain 16.191 g of crystals of the synisomer of sodium3-acetocymethyl-7-[2-(2-amino-4-thiazolyl)-2-methoxyiminoacetamido]-ceph-3-eme-4-carboxylate.

By operating while avoiding contact with atmospheric humidity, thephysical properties of the product were: 0.2% of water (Karl Fischer);0.1% methanol and; 0.45% ethanol with the latter two values beingdetermined by vapor phase chromatography.

Analysis: C₁₆ H₁₆ O₇ N₅ S₂ Na; molecular weight=477.5: Calculated: %C40.24, %H, 3.38, %N, 14.67, %S, 13.43, %Na, 4.8 Found: %C, 39.9, %H,3.5, %N, 14.5, %S, 13.1, %Na, 4.8.

The product reehydrated when standing in air. The x-ray spectrum (DebyeScherrer) confirmed the crystalline nature of the product. Isopropanolin place of ethanol was equally useful for the crystallization.

EXAMPLE 24 cristallize syn isomer of sodium3-acetoxymethyl-7-[2-(2-amino-4-thiazolyl)-2-methoxyiminoacetamido]-ceph-3-eme-4-carboxylateSTEP A: solvate of formic acid and syn isomer of3-acetoxymethyl-7-[2-(2-amino-4-thiazolyl)-2-methoxyiminoacetamido]-ceph-3-eme-4-carboxylate

87.2 g of the diethylamine salt of3-acetoxymethyl-7-[2-(2-tritylamino-4-thiazolyl)-2-methoxyiminoacetamido]-ceph-3-eme-4-carboxylicacid of Example 5 was added in small amounts with stirring to a mixtureof 220 ml of formic acid and 220 ml water and the mixture was stirred at50° C. for 30 minutes and was then cooled. The mixture was filtered toremove 30.1 g of triphenyl carbinol and the filtrate was poured into 450ml of water. The mixture was treated with carbon and filtered and thefilter was evaporated at 40° C. under reduced pressure until aprecipitate formed. 200 ml of anhydrous ethanol were added and themixture was cooled with ice and was filtered. The precipitate was washedwith ethanol and with ether and was dried under reduced pressure toobtain 31.1 g of a solvate of formic acid and the syn isomer of3-acetoxymethyl-7-[2-(2-amino-4-thiazolyl)-2-methoxyiminoacetamido]-ceph-3-eme-4-carboxylicacid.

Analysis: C₁₆ H₁₇ N₅ O₇ S₂. HCOOH . H₂ O; molecular weight=541.5Calculated: % C 39.3, % H 4.08, % N 13.48, % S 12.34, % H₂ O, 3.46.Found: 39.2, 4.1, 13.2, 12.8, %H₂ O, 4.15.

STEP B: crystallize sodium salt of the syn isomer of3-acetoxymethyl-7-[2-(2-amino-4-thiazolyl)-2-methoxyiminoacetamido]-ceph-3-eme-4-carboxylicacid

A solution of 15 g of the freshly prepared solvate of Step A in 75 ml ofethanol was treated with 4.5 g of potassium acetate and 3 g of activecarbon and was then filtered. 5 ml of isopropanol were added withstirring to the filtrate and after standing at 0° C. for 15 hours, themixture was filtered. The crystals were washed with ethanol and etherand dried for 2 hours at 50° C. under reduced pressure to obtain 7.95 gof the syn isomer of sodium3-acetoxymethyl-7[2-(2-amino-4-thiazolyl)-2-methoxyiminoacetamido]-ceph-3-eme-4-carboxylate.The product stood briefly in free and was then analyzed.

Analysis: C₁₆ H₁₆ N₅ NaO₇ S₂. H₂ O; molecular weight=495.5: Calculated:%C, 38.78, %H, 3.66, %N, 14.14, % Na, 4.64, %S, 12.94. Found: %C, 38.6,%H, 3.7, %N, 13.8, %Na, 4.6, %S, 13.2.

EXAMPLE 25 amorphous syn isomer of sodium3-acetoxymethyl-7-[2-(2-amino-4-thiazolyl)-2-methoxyiminoacetamido]-ceph-3-eme-4-carboxylateSTEP A: solvate of ethanol and syn isomer of3-acetoxymethyl-7-[2-(2-amino-4-thiazolyl)-2-methoxyiminoacetamido]-ceph-3-eme-4-carboxylicacid

A solution of 52 g of the solvate of formic acid obtained in Step A ofExample 24 in 3 liters of 96% ethanol and 350 ml of water wasconcentrated under reduced pressure to obtain a value of about 300 mland the solvate began to crystallize during the concentration. Themixture was cooled for an hour in an ice bath and was filtered. Theprecipitate was washed with a little ethanol and was dried at roomtemperature under reduced pressure in the presence of concentratedsulfuric acid to obtain 44 g of the solvate of ethanol and the synisomer of3-acetoxymethyl-7-[2-(2-amino-4-thiazolyl)-2-methoxyiminoacetamido]-ceph-3-eme-4-carboxylicacid.

Analysis: C₁₆ H₁₇ N₅ O₇ S₂ . 0.8 C₂ H₅ OH; molecular weight=492.3:Calculated: %C, 42.94, %H, 4.46, %N, 14.23, %S, 13.02. Found: %C, 43.0,%H, 4.4, %N, 14.1, %S, 12.9.

STEP B: amorphous sodium salt

A mixture of 3 g of the solvate of Step A in 60 ml of water was cooledto 0° C. and a solution of 0.504 g of sodium bicarbonate in 6 ml ofwater was added thereto. The neutral solution was filtered andlyophilysed immediated. After standing briefly in free air the productwas analyzed.

Analysis: C₁₆ H₁₆ N₅ NaO₇ S₂ 1.5H₂ O; molecular weight=504.47:Calculated: %C 38.09, %H, 3.8, %N, 13.85. Found: %C, 38.2, %H, 3.9, %N,13.6.

EXAMPLE 26 cristallize syn isomer of sodium3-acetoxymethyl-7-[2-(2-amino-4-thiazolyl)-2-methoxyiminoacetamido]-ceph-3-eme-4-carboxylate

0.25 ml of n-butanol were slowly added with stirring to a solution of0.5 g of the amorphous salt of Example 25 in 2 ml of methanol and themixture was held at 6° C. for 48 hours. The crystals were washed with alittle cold methanol and was dried for 3 hours at 40° C. under reducedpressure in the presence of concentrated sulfuric acid to obtain 0.2 gof the above crystalline sodium salt. The product was analyzed afterbriefly being exposed to air.

Analysis: C₁₆ H₁₆ N₅ NaO₇ S₂ 1.5H₂); molecular weight=504.4: Calculated:%C, 38.09, %H, 3.8, %N, 13.88, %O, 29.96. Found: %C, 38.4, %H, 3.8, %N,13.8, %O, 27.1.

Under analogous operating conditions, slightly different crystallineforms were obtained containing 0.1 mole of methanol and 0.5 or 1 mole ofwater. The x-ray spectra (Debye Scherrer of the products confirmed therenature.

EXAMPLE 27 crystalline syn isomer of sodium3-acetoxymethyl-7-[2-(2-amino-4-thiazolyl)-2-methoxyiminoacetamido]-ceph-3-eme-4-carboxylate

A mixture of 4.95 g of the product of Example 22 in 5 ml of ethanol wasstirred n an ice bath and 10 ml of a molar solution of aqueous sodiumbicarbonate were added. After dissolution, 15 ml of ethanol were addedand the mixture was evaporated to dryness under reduced pressure at 30°C. The residue was taken up in 15 ml of methanol and crystallization wasinduced. The mixture stood overnight in the refrigerator to obtain 3.407g of the sodium salt identical to the product of Example 23.

EXAMPLE 28 syn isomer of7-[2-(2-amino-4-thiazolyl)-2-hydroxyiminoacetamido]-3-acetoxymethyl-ceph-3-eme-4-carboxylicacid STEP A: syn isomer of ethyl2-(2-amino-4-thiazolyl)2-hydroxyimino-acetate

2 g of ethyl 4-chloro-2-hydroxyimino-acetyl acetate were added over 5minute to a solution of 0.8 g of thiourea in 2.4 ml of ethanol and 4.8ml of water and the mixture was stirred for one hour at roomtemperature. The majority of the ethanol was distilled off under apartial pressure and the mixture was adjusted to a pH of 6 by additionof solid sodium bicarbonate. The mixture was iced and vacuum filteredand the recovered product was washed with water and dried under reducedpressure at 40° C. to obtain 1.32 of ethyl2-(2-amino-4-thiazolyl)-2-hydroxyimino-acetate melting at 232° C.

Analysis: C₅ H₉ O₃ N₃ S: Calculated: %C, 39.06, %H, 4.21, %N, 19.52, %S,14.9. Found: %C, 38.9, %H, 4.4, %N, 19.7, %S, 14.6.

RMN (DMSO, 60 MHz): (a) triplet centered about 1.25 ppm J=7 Hz; (b)quadruplet centered about 4.27 ppm J=7 Hz; (c) singulet at 6.83 ppm; (d)singulet at 7.11 ppm; (e) singulet at 11.4 ppm. ##STR39##

STEP B: syn isomer of 2-(2-amino-4-thiazolyl)-2-hydroxyimino-acetic acid

A mixture of 21.5 g of the product of Step A, 200 ml of absolute ethanoland 55 ml of 2N sodium hydroxide was stirred for 45 minutes on a waterbath at 45° C. and was then placed in an ice water bath. The pH wasadjusted to 6 with acetic acid and a precipitation was observed and themixture was vacuum filtered. The recovered product was rinsed with a 1-1ethanol-water mixture and then with ether and dried to obtain 16.9 g ofsyn isomer of 2-(2-amino-4-thiazolyl)-2-hydroxyimino-acetic acid with anRf=0.05 (eluant-70-20-10 ethyl acetate-ethanol-water).

STEP C: sodium salt of syn isomer of2-(2-tritylamino-4-thiazolyl)-2-tritylhydroxyimino-acetic acid

A mixture of 16.9 g of the acid of Step B, 50 ml of dimethylformanideand 42 ml of triethylamine was stirred at room temperature for 15minutes to effect total dissolution and the mixture was cooled to -20°C. to partially crystallize the triethylamine salt. A mixture of 54 g oftrityl chloride in 100 ml of chloroform was added over 15 minute at -20°C. to the mixture which was then stirred for an hour during which thetemperature returned to room temperature. The mixture was poured into200 ml of water containing 40 ml of 2N hydrochloric acid. The mixturewas decanted and the organic phase was washed twice with 200 ml ofwater, was dried and vacuum filtered. The filtrate was evaporated todryness under reduced pressure and the residue was taken up in ethylacetate 100 ml of a saturated aqueous sodium bicarbonate solution wasadded thereto and the mixture was stirred and decanted. Crystallizationoccured while icing for 30 minutes and the mixture was vacuum filtered.The recovered product was rinsed with ethyl acetate to obtain 27 g ofsodium 2-(2-tritylamino-4-thiazolyl)-2-tritylhydroxylimino-acetate witha Rf=0.33 (ether).

STEP D: syn isomer of tert.-butyl7-[2-(2-tritylamino-4-thiazolyl-2-tritylhydroxyimino-acetamido]-3-acetoxymethylceph-3-eme-4-carboxylate

A mixture of 17.2 g of the sodium salt of Step C, 170 ml of chloroformand 170 ml of N hydrochloric acid was decanted and the organic phase waswashed 5 times with water, dried and vacuum filtered. The filtrate wasevaporated to dryness and the residue was taken up in 170 ml ofmethylene chloride. 2.8 g of dicyclohexyldicarbondimide were addedthereto and the mixture was stirred for an hour after which it wasvacuum filtered to remove 1.9 g of dicyclohexylurea. 3.66 g oftert.-butyl 7-amino-3-acetoxymethyl-ceph-3-eme-4-carboxylate were addedto the filtrate and the mixture was stirred for 2 hours at roomtemperature. The mixture was washed with N hydrochloric acid, withwater, with an aqueous 5% sodium bicarbonate solution and finally withwater. The organic phase was dried and vacuum filtered and the filtratewas evaporated to dryness. The residue was taken up in methylenechloride and the solution was chrometographed over silica gel. Elutionwas effected with methylene chloride containing 5% ether and thefractions with an Rf=0.78 in ether were combined and evaporated todryness under reduced pressure. The residue was taken up isopropyl etherand effloresced. The mixture was vacuum filtered and the product wasrinsed with isoproyl ether to obtain 5.8 g of the syn isomer oftert.-butyl7-[2-(2-tritylamino-4-thiazolyl)2-tritylhydroxyimino-acetamido]-3-acetoxymethyl-ceph-3-eme-4-carboxylate.

Analysis: C₅₇ H₅₁ O₇ N₅ S₂ Calculated: %C, 69.7, %H, 5.2, %N, 7.1, %S,6.5 Found: 70.4 5.6 6.5 5.9

RMN (CDCl₃ 60 MHz): (a) singulet at 1.55 ppm; (b) singulet at 2.06 ppm;(c) singulet at 6.45 ppm; (d) singulet at 7.31 ppm. ##STR40##

STEP E: syn isomer of7-[2-(2-amino-4-thiazolyl)-2-hydroxyimino-acetamido]-3-acetoxymethyl-ceph-3-eme-4-carboxylicacid

A mixture of 1 g of the product of Step D in 3 ml of trifluoroaceticacid was stirred at room temperature for 30 minutes and then 30 ml ofisopropyl ether were added thereto. The mixture was vacuum filtered torecover the precipitated salt which was rinsed with isopropyl ether toobtain 0.652 g of the salt of trifluoroacetic acid and7-[2-(2-amino-4-thiazolyl)-2-tritylhydroxyimino-acetamido]-3-acetoxymethyl-ceph-3-eme-4-carboxylicacid. The said salt was dissolved it 6 ml of tetrahydrofuran and 3 ml of50% aqueous formic acid were added thereto. The mixture was stirred at50° C. for 15 minutes and was evaporated to dryness. The residue wastaken up in ether and the mixture was filtered. The product was rinsedwith ether to obtain 0.441 g of the formate of the product which wastriturated with 2 ml of water containing 3 drops of pyridine (pH≃6). Themixture was vacuum filtered and the product was rinsed with water anddried to obtain 0.136 mg of the syn isomer of7-[2-(2-amino-4-thiazolyl)-2-hydroxyimino-acetamido]-3-acetoxymethyl-ceph-3-eme-4-carboxylicacid. The filtrate was evaporated to dryness and the residue was takenup in ether. The mixture was vacuum filtered and the product was rinsedwith ethanol to obtain another 0.04 of the said syn isomer.

RMN (DMSO, 60 MHz): (a) singulet at 2.01 ppm; (b) singulet at 6.67 ppm;(c) singulet at 7.08 ppm and (d) singulet at 11.3 ppm. ##STR41##

EXAMPLE 29 syn isomer of7-[2-(2-amino-4-thiazolyl)2-hydroxyiminoacetamido]-3-acetoxymethyl-ceph-3-eme-4-carboxylicacid STEP A: syn isomer of ethyl2-(2-tritylamino-4-thiazolyl)-2-hydroxyimino-acetate

32 ml of triethylamine were added to a mixture of 43.2 g of the synisomer of ethyl 2-(2-amino-4-thiazolyl)-2-hydroxylimino-acetate in 120ml of dry dimethylformamide cooled to -35° C. and then 60 g of tritylchloride were added thereto in small fractions over 30 minutes. Thetemperature returned to room temperature during which total dissolutionoccured and the mixture was heated to 30° C. for an hour. The mixturewas poured into 1.2 liters of iced water containing 40 ml of 22° Behydrochloric acid and the mixture was stirred on an ice bath and wasthen vacuum filtered. The precipitate was rinsed with N hydrochloricacid and was empasted with ether to obtain 69.3 g of a hydrochloridesalt. The product was dissolved in 5 volumes of methanol to which wasadded 120% of triethylamine and 5 volumes of water were added tocrystalize the syn isomer of ethyl2-(2-tritylamino-4-thiazolyl)-2-hydroxylimino-acetate.

Analysis: C₂₆ H₂₃ O₃ N₃ S. 1/4H₂ O: Calculated: %C, 67.6, %H, 5.1, %N,9.1, %S, 6.9. Found: %C, 67.5, %H, 5.1, %N, 8.8, %S, 6.8.

RMN (CDCl₃, 60 MHz): (a) triplet centered about 1.31 ppm, J=7 Hz; (b)quadruplet centered about 4.37 ppm, J=7 Hz; (c) singulet at 6.37 ppm;and (d) singulet at 7.28 ppm. ##STR42##

STEP B: syn isomer of ethyl2(2-tritylamino-4-thiazolyl)-2-tetrahydrophyranyloxyimino-acetate

2.4 g of p-toluene sulfonic acid were added to a mixture of 5.6 g of theproduct of Step A in 56 ml of redistilled dihydropyran in an ice bathand the mixture was then stirred for an hour while the temperaturereturned to room temperature. The mixture was poured into a mixture of100 ml of benzene, 100 ml of water and 2 ml of triethylamine and theorganic phase was decanted, was washed with water, dried and vacuumfiltered. The filter was rinsed with benzene and the filtrate wasevaporated to dryness. The residue was taken up in isoproyl andcrystallization was induced. The mixture was stored overnight in arefrigerator and was then vacuum filtered. The product was rinsed withisopropyl ether to obtain 4.42 g of the syn isomer of ethyl2-(2-tritylamino-4-thiazolyl)-2-tetrahydropyranyloxyimino-acetatemelting at 184° C.

Analysis: p-toluene sulfonic acid salt C₃₈ H₃₉ O₇ N₃ S₂ : Calculated:%C, 63.9, %H, 5.5, %N, 5.9, %S, 9.0. Found: %,C 63.7, %H, 5.5, %N, 5.8,%S, 8.9.

RMN (CDCl₃, 60 MHz): (a) triplet centered about 1.36 ppm; (b) quadrupletcentered about 4.39 ppm; (c) singulet at 6.60 ppm; (d) sigulet at 6.91ppm; and (e) sigulet at 7.28 ppm. ##STR43##

STEP C: syn isomer of2-(2-tritylamino-4-thiazolyl)-2-tetrahydropyranyloxyimino-acetic acid

A mixture of 4.56 g of the product of Step B, 45 ml of dioxane and 8.4ml of 2N sodium hydroxide was refluxed for 90 minutes and was thencooled on an ice-water bath. The mixture was vacuum filtered and therecovered product was rinsed with aqueous dioxane and then with ether ofobtain 4.66 g of a sodium salt. The latter was dissolved in 50 ml ofdioxane and the solution was acidified with formic acid to a pH of 5.The addition of 90 ml of water caused precipitation of the syn isomer of2-(2-tritylamino-4-thiazolyl)-2-tetrahydropyranyloxyimino-acetic acidmelting at 180° C.

RMN (CDCl₃, 60 MHz): 6.69 ppm (portion of thiazolyl ring) and 7.31(aromatic).

STEP D: syn isomer of tert.-butyl7-[2-(2-tritylamino-4-thiazolyl)-2-tetrahydropyranyloxyimino-acetamido]-3-acetoxymethyl-ceph-3-eme-4-carboxylate

A mixture of 0.362 g of the product of Step C, 0.244 g of tert.-butyl7-amino-3-acetoxymethyl-ceph-3-eme-4-carboxylate, 0.280 g ofdicyclohexylcarbodiimide and 4 ml of dry chloroform was stirred for 2hour at room temperature and the mixture was vacuum filtered to removedicyclohexylurea which was rinsed with chloroform. The filtrate wasevaporated to dryness under reduced pressure and the residue wasdissolved in 1 ml of ether. The solution was chromatographed over silicagel and was eluted with ether. The fractions with an Rf=0.38 werecombined and evaporated to dryness under reduced pressure. The residuewas taken up in isopropyl ether and effloresced. The mixture was vacuumfiltered and the recovered product was rinsed with isopropyl ether toobtain 0.184 g of the syn isomer of tert.-butyl7-[2-(2-tritylamino-4thiazolyl)-2-tetrahydropyranyloxyimino-acetamido]-3-acetoxymethyl-ceph-3-eme-4-carboxylate.

Analysis: C₄₃ H₄₅ O₈ N₅ S₂ : Calculated: %C, 62.7, %H, 5.5, %N, 8.5, %S,7.8. Found: %C, 62.8, %H, 5.9, %N, 8.1, %S, 7.5.

RMN (CDCl₃, 60 MHz): (a) 1.53 ppm; (b) 2.07 ppm; (c) 5.46 ppm; (d) 6.76ppm; and (e) 7.28 ppm. ##STR44##

STEP E: syn isomer of7-[2-(2-amino-4-thiazolyl)-2-hydroxyimino-acetamido]-3-acetoxymethyl-ceph-3-eme-4-carboxylicacid

A mixture of 638 mg of the product of Step D in 1.8 ml oftrifluoroacetic acid was stirred for 15 minutes at room temperature and18 ml of isopropyl ether was added thereto. The mixture was vacuumfiltered to obtain 404 mg of product which was added to 2 ml of 50%aqueous formic acid. The mixture was stirred for 15 minutes at 50° C.and was then evaporated to dryness at 30° C. under reduced pressure. Theresidue was taken up in 1 ml of ethanol and after adding a drop ofpyridine thereto, the mixture was vacuum filtered to obtain the synisomer of7-[2-(2-amino-4-thiazolyl)-2-hydroxyiminoacetamido]-3-acetoxymethyl-ceph-3-eme-4-carboxylicacid which was identical to the product of Example 28.

EXAMPLE 30 syn isomer of7-[2-(2-tritylamino-4-thiazolyl)-2-tritylhydroxyimino-acetamido]-3-acetoxymethyl-ceph-3-eme-4-carboxylicacid STEP A: syn isomer of ethyl2-(2-tritylamino-4-thiazolyl)-2-tritylhydroxyimino-acetate

1.5 ml of triethylamine were added to a solution of 1.08 g of theproduct of Step A of Example 28 in 8 ml of chloroform and then asolution of 3 g of trityl chloride in 6 ml of chloroform was addedthereto at 5° C. over 25 minutes. The mixture was stirred for an hour atroom temperature and the mixture was washed with 18 ml of water, then 8ml of N hydrochloric acid and then 3 times with 20 ml of water. Themixture was dried and evaporated to dryness and the residue wascrystallized from isopropanol to obtain 2.3 g of the syn isomer of ethyl2-(2-tritylamino-4-thiazolyl)-2-tritylhydroxyimino-acetate melting at140° C.

STEP B: sodium salt of syn isomer of2-(2-tritylamino-4-thiazolyl)-2-tritylhydroxyimino-acetic acid

0.7 g of the product of Step A was dissolved in 3.5 ml of hot dioxaneand 1 ml of 2N sodium hydroxide solution was added thereto dropwise withstirring at 110° C. The mixture was stirred for 110 minutes at refluxwith stirring and was then cooled and vacuum filtered to obtain thesodium salt of the syn isomer of2-(2-tritylamino-4-thiazolyl)-2-tritylhydroxyimino-acetic acid which wasidentical to Step C of Example 28.

STEP C: syn isomer of7-[2-(2-tritylamino-4-thiazolyl)-2-tritylhydroxyimino-acetamido]-3-acetoxymethyl-ceph-3-eme-4-carboxylicacid

50 ml of N hydrochloric acid were added with stirring to a suspension of5.12 g of the salt of Step B in 50 ml of chloroform and the organicphase was decanted and was washed 3 times with 50 ml of water. Themixture was dried and was vacuum filtered and the filtrate wasevaporated to dryness to obtain the syn isomer of2-(2-tritylamino-4-thiazolyl)-2-tritylhydroxyimino-acetic acid. The saidproduct was dissolved in 50 ml of methylene chloride and 1.6 g ofdicyclohexylcarbodiimide were added thereto. The mixture was stirred foran hour at room temperature and was vacuum filtered to removedicyclohexylurea. The filtrate was cooled to -10° C. and a solution of1.1 g of 7-amino-cephalosporanic acid in 10 ml of methylene chloride and1.2 ml of triethylamine was added thereto. The mixture was stirred for 2hour at room temperature and 50 ml of N hydrochloric acid were addedwith stirring. The organic phase was decanted, was washed 3 times with50 ml of water, was dried and vacuum filtered. The filtrate wasevaporated to dryness and the residue was efloresced with ethanol. Themixture was filtered to obtain 2.36 of raw product which was reactedwith diethylamine in ether to obtain the diethylamine salt of7-[2-(2-tritylamino-4-thiazolyl)-2-tritylhydroxyimino-acetamido]-3-acetoxymethyl-ceph-3-eme-4-carboxylic acid. The said salt was dissolved inmethylene chloride and N hydrochloric acid was added thereto until thepH was acid. The solution was washed with water, dried and evaporated todryness to obtain 0.75 of synisomor of 7-[2(2-tritylamino-4-thiazolyl)-2-tritylhydroxyimino-acetamido]-3-acetoxymethyl-ceph-3-eme-4-carboxylicacid.

EXAMPLE 317-[2-(2-amino-4-thiazolyl)-2-hydroxyimino-acetamido]-3-acetoxymethyl-ceph-3-eme-4-carboxylicacid.

A solution of the product of Example 30 in 5 ml of 50% aqueous formicacid was stirred at 50° C. for 15 minutes and was then evaporated todryness. The residue was taken up in ether and the mixture was vacuumfiltered. The product was rinsed with ether to obtain the syn isomer of7-[2-(2-amino-4-thiazolyl)-2-hydroxylmino-acetamido]-3-acetoxymethyl-ceph-3-eme-4-carboxylicacid in the form of its formate. The latter was triturated with 3 ml ofwater containing a few drops of pyridine (pH≃6). The mixture was vacuumfiltered and the recovered product was rinsed with water and dried toobtain the said acid which was identical to the product of Example 28.

EXAMPLE 32 sodium salt of syn isomer of7-[2-(2-amino-4-thiazolyl)-2-hydroxyimino-acetamido]-3-acetoxymethyl-ceph-3-eme-4-carboxylicacid

A solution of 0.613 g of the product of Example 28 in 2 ml of distilledwater and 2 ml of a M solution sodium acetate in methanol was admixedwith 60 mg of activated carbon and the mixture was vacuum filtered. Thefilter was rinsed with 2 ml of methanol and the filtrate was evaporatedto dryness at 30° C. under reduced pressure. The residue was taken up inether and was filtered. The product was dried to obtain 0.432 g of thesodium salt of the syn isomer of7-[2-(2-amino-4-thiazolyl)2-hydroxyimino-acetamido]-3-acetoxymethyl-ceph-3-eme-4-carboxylicacid.

EXAMPLE 33 Crystalline sodium salt of the syn isomer of7-(2-amino-4-thiazolyl)-2-hydroxyimino-acetamido]-3-acetoxymethyl-ceph-3-eme-4-carboxylicacid

8 ml of a molar solution of sodium acetate in methanol were added to asuspension of 1.78 g of the acid of Example 28 in 8 ml of methanol andthe sodium salt immediately crystallized. The mixture was stirred for 15minutes to obtain complete crystallization and was vacuum filtered. Theproduct was washed with ethanol and then with ether to obtain 1.53 g ofthe crystalline sodium salt of the syn isomer of7-[2-(2-amino-4-thiazolyl)-2-hydroxyimino-acetamido]-3-acetoxymethyl-ceph-3-eme-4-carboxylicacid as white crystals.

Analysis: C₁₅ H₁₄ O₇ N₅ S₂ Na: Calculated: %C, 38.88, %H, 3.04, %N,15.11, %S, 13.84, %Na, 4.96. Found: %C, 38.8, %H, 3.1, %N, 15.1, %S,13.8, %Na, 4.85.

Infrared Spectrum (Nujol): >C=0 at 1753, 1724 and 1683 cm⁻¹ ; --NH andOH at 3597 cm⁻¹.

RMN (DMSO, 60 MHz): singulet at 2.01 ppm ##STR45## singulet at 6.65 ppm(proton of thiazole ring); and singulet at 7.16 ppm (amine NH₂).

EXAMPLE 34

An injectable solution was prepared with 500 mg of3-acetoxymethyl-7-[2-(2-amino-4-thiazolyl)-2-hydroxyiminoacetamido]-ceph-3-eme-4-carboxylicacid or3-acetoxymethyl-7-[2-(2-amino-4-thiazolyl)-2-methoxyimino-acetamido]-ceph-3eme-4-carboxylicacid or the syn isomer of7-[2-(2-amino-4-thiazolyl)-2-hydroxyimino-acetamido]-3-acetoxymethyl-ceph-3-eme-4-carboxylicacid and sufficient sterile water to make a final solution of 5 ml. Asimilar solution was prepared with 500 mg of sodium3-acetoxymethyl-7-[2-(2-amino-4-thiazolyl)-2-methoxyimino-acetamido]-ceph-3-eme-4-carboxylicacid or3-acetoxymethyl-7-[2-(2-amino-4-thiazolyl)-2-(1-methylethoxyimino)-acetamido]-ceph-3-eme-4-carboxylicacid.

Gelules were prepared with 250 mg of 3-acetoxymethyl7-[2-(2-amino-4-thiazolyl)-2-methoxyimino-acetamido]-ceph-3-eme-4-carboxylicacid or sodium3-acetoxymethyl-7-[2-(2-amino-4-thiazolyl)-2-methoxyimino-acetamido]-ceph-3-eme-4-carboxylateor3-acetoxymethyl-7-[2-(2-amino-4-thiazolyl)-2-(1-methylethoxyimino)-acetamido]-ceph-3-eme-4-carboxylicacid or the syn isomer of7-[2-(2-amino-4-thiazolyl)-2-hydroxyimino-acetamido]-3-acetoxymethyl-ceph-3-eme-4-carboxylicacid and sufficient excipient to obtain a final weight or 400 mg.

PHARMACOLOGICAL DATA A. In Vitro Activity

The method used was a dilution of a liquid medium where a series oftubes received the same quantity of a sterile nutritive media andincreasing doses of the test compounds were placed therein. Then eachtube was seeded with a bacterial strain and was incubated for 24 to 48hours at 37° C. in an oven. The increasing inhibition was determined bytransillumination to determine the minimum inhibiting concentration (MICin μg/ml) and the results are reported in the follow Tables.

Product A is the anti isomer of3-acetoxymethyl-7-[2-(2-amino-4-thiazolyl)-2-methoxyimino-acetamido]-ceph-3-eme-4-carboxylicacid.

    __________________________________________________________________________    Anti isomer of 3-acetoxymethyl-7-[2-(2-amino-4-thiazolyl)-2-hydroxy-          iminoacetamido]-ceph-3-eme-4-carboxylic acid                                                                M.I.C. in μg/ml                                      STRAINS               24 H                                                                              48 H                                        __________________________________________________________________________            Staphylococcus aureus UC 1061 Pen-Sensible                                                          5   5                                                   Staphylococcus aureus UC 1128 Pen-Resistant                                                         10  10                                                  Staphylococcus aureus Exp. No 54146                                                                 5   10                                                  Streptococcus pyogenes A 561                                                                        0,5 0,5                                                 Bacillus subtilis ATCC 6633                                                                         2   5                                                   Escherichia Coli ST UC 1020                                                                         3   3                                                   Escherichia Coli RT UC 1261                                                                         1   1                                                   Escherichia Coli Exp. TO.sub.26 B.sub.6                                                             2   2                                                   Escherichia Coli RG R55 123D                                                                        5   10                                                  Klebsiella pneumoniae Exp. 54145                                                                    0,6 0,6                                                 Klebsiella pneumoniae 2536 R                                                                        20  40                                                  Proteus mirabilis (indol-) A 235                                                                    2   2                                                   Salmonella typhimurium 420                                                                          2   2                                                   Enterobacter cloacae 681                                                                            40  40                                          __________________________________________________________________________                        Product of                                                                             Product of                                                           Example 4                                                                              Example 7                                                                              Product A                                                   M.I.C. in μg/ml                                                                     M.I.C. in μg/ml                                                                     M.I.C. in μg/ml                      STRAINS             24 H 48 H                                                                              24 H                                                                              48 H 24 H 48 H                               __________________________________________________________________________    Staphylococcus aureus ATCC 5 638 Pen-                                                             1    1   1   2    40   40                                 Sensible = UC 1061                                                            Staphylococcus aureus UC 1 128 Pen-                                                               2    2   2   3    >40  >40                                Resistant                                                                     Staphylococcus aureus exp. no 54 146                                                              3    3   2   2    40   40                                 Streptococcus pyogenes A 561                                                                      0,02 0,05                                                                              0,01                                                                              0,01 0,4  0,4                                Streptococcus faecalis 5 432                                                                      2    2   1   3    >40  >40                                Streptococcus faecalic 99 F 74                                                                    2    3   3   10   >40  >40                                Bacillus subtilis ATCC 6 633                                                                      0,2  1   1   2    20   20                                 Escherichia Coli Sensible Tetracycline                                                            0,2  0,2 0,1 0,1  20   40                                 ATCC 9 637 = UC 1020                                                          Escherichia Coli Resistant Tetracycline                                                           0,02 0,02                                                                              ≦0,02                                                                      ≦0,02                                                                       2    2                                  ATCC 11 303 = UC 1261                                                         Escherichia Coli Exp. TO.sub.26 B.sub.6                                                           0,1  0,1 0,05                                                                              0,05 5    5                                  Escherichia Coli Resistant Gentamicine,                                                           0,6  0,6-1                                                                             0,1 0,1  10   10                                 Tobramycine R 55 123 D                                                        Klebsiella pneumoniae Exp. 52 145                                                                 0,02 0,02                                                                              ≦0,02                                                                      ≦0,02                                                                       1    1                                  Klebsiella pneumoniae 2 536 Resistant                                                             0,6  0,6 0,2 0,2  >40  >40                                Gentamicine                                                                   Proteus mirabilis (indol-) A 235                                                                  0,02 0,02                                                                              ≦0,02                                                                      ≦0,02                                                                       2    2                                  Proteus vulgaris (indol+) A 232                                                                   2    40  0,2 0,6  >40  >40                                Salmonella typhimurium 420                                                                        0,2  0,2 0,1 0,1  20   20                                 Providencia Du 48   5    5   1   2    40   >40                                Pseudomonas 3 935 Exp. SG                                                                         10   40  10  20   >40  >40                                Serratia Resistant Gentamicine 2 532                                                              1    1   1   1    >40  >40                                Enterobacter cloacae 681                                                                          40   40                                                   Pseudomonas 8951 RGT                                                                              40   >40                                                  Staphylococcus Smith                                                                              1    1                                                    Escherichia Coli Galle 0,1                                                                        0,2                                                       Escherichia Coli E.sub.6                                                                          0,05 0,05                                                 Escherichia Coli T 25 575                                                                         0,05 0,05                                                 Escherichia Coli T 96 875                                                                         0,05 0,05                                                 __________________________________________________________________________                                 Product of                                                                    Example 9                                                                     M.I.C. in μg/ml                                        STRAINS             24 H                                                                              48 H                                         __________________________________________________________________________             Staphylococcus aureus ATCC 5 638 Pen-                                                             1   2                                                     Sensible                                                                      Staphylococcus aureus UC 1 128 Pen-                                                               2   2                                                     Resistant                                                                     Staphylococcus aureus exp. no 54 146                                                              2   2                                                     Staphylococcus aureus Co 15 Resistant                                                             10  20                                                    Cephalexine                                                                   Streptococcus pyogenes A 561                                                                      0,02                                                                              0,02                                                  Streptococcus faecalis 5 432                                                                      2   10                                                    Bacillus subtilis ATCC 6 633                                                                      1   1                                                     Escherichia Coli Sensible Tetracycline                                                            1   1                                                     ATCC 9 637                                                                    Escherichia Coli Resistant Tetracycline                                                           0,1 0,1                                                   ATCC 11 303                                                                   Escherichia Coli Exp. TO.sub.26 B.sub.6                                                           1   1                                                     Escherichia Coli Resistant Gentamicine,                                                           1   1                                                     Tobramycine R 55 123 D                                                        Klebsiella pneumoniae Exp. 52 145                                                                 0,05                                                                              0,1                                                   Klebsiella pneumoniae 2 536 Resistant                                                             2   5                                                     Gentamicine                                                                   Proteus mirabilis (indol-) A 235                                                                  0,2 0,2                                                   Proteus vulgaris (indol+) A 232                                                                   2   10                                                    Salmonella typhimurium 420                                                                        1   10                                                    Providencia Du 48   3   5                                                     Pseudomonas 3 935 Exp. SG                                                                         20  20                                                    Serratia Resistant Gentamicine 2 532                                                              0,4 1                                            __________________________________________________________________________                             Product of                                                                             Product of                                                           Example 11                                                                             Example 15                                                           M.I.C. in μg/ml                                                                     M.I.C. in μg/ml                          STRAINS                  24 H 48 H                                                                              24 H 48 H                                   __________________________________________________________________________    Staphylococcus aureus ATCC 5 638 Pen-                                                                  2    2   20   20                                     Sensible                                                                      Staphylococcus aureus UC 1 128 Pen-                                                                    2    3   40   40                                     Resistant                                                                     Staphylococcus aureus exp. no 54 146                                                                   2    2   40   40                                     Streptococcus pyogenes A 561                                                                           0,02 0,02                                                                              0,4  0,4                                    Streptococcus faecalis 5 432                                                                           2    10  >100 >100                                   Streptococcus faecalic 99 F 74                                                                         2    20  >100 >100                                   Bacillus subtilis ATCC 6 633                                                                           0,4  0,4 10   10                                     Escherichia Coli Sensible Tetracycline                                                                 1    1   >100 >100                                   ATCC 9 637                                                                    Escherichia Coli Resistant Tetracycline                                                                ≦0.2                                                                        0.2 10   10                                     ATCC 11 303                                                                   Escherichia Coli Exp. TO.sub.26 B.sub.6                                                                0.4  0.4 20   40                                     Escherichia Coli Resistant Gentamicine,                                                                0.6  0.6 40   40                                     Tobramycine R 55 123 D                                                        Klebsiella pneumoniae Exp. 52 145                                                                      0.05 5   20   20                                     Klebsiella pneumoniae 2 536 Resistant                                                                  2    2   >100 >100                                   Gentamicine                                                                   Proteus mirabilis (indol-) A 235                                                                       0.05 5   5    5                                      Proteus vulgaris (indol+) A 232                                                                        1    2   >100 >100                                   Salmonella typhimurium 420                                                                             1    1   100  100                                    Providencia Du 48        5    5   100  >100                                   Pseudomonas 3 935 Exp. SG                                                                              20   40  >100 >100                                   Serratia Resistant Gentamicine 2 532                                                                   2    2   >100 >100                                   Enterobacter cloacae 681 40   40  100  100                                    __________________________________________________________________________                             Product of                                                                             Product of                                                           Example 13                                                                             Example 17                                                           M.I.C. in μg/ml                                                                     M.I.C. in μg/ml                          STRAIN                   24 H 48 H                                                                              24 H 48 H                                   __________________________________________________________________________    Staphylococcus aureus ATCC 5 638 Pen-                                                                  2    5   20   20                                     Sensible                                                                      Staphylococcus aureus UC 1 128 Pen-                                                                    3    5   40   40                                     Resistant                                                                     Staphylococcus aureus exp. no 54 146                                                                   2    5   40   40                                     Streptococcus pyogenes A 561                                                                           0,02 0,02                                                                              1    1                                      Streptococcus faecalis 5 432                                                                           5    40  >100 >100                                   Streptococcus faecalic 99 F 74                                                                         5    20  >100 >100                                   Bacillus subtilis ATCC 6 633                                                                           0,5  2   20   >100                                   Escherichia Coli Sensible Tetracycline                                                                 2    2   >100 >100                                   ATCC 9 637                                                                    Escherichia Coli Resistant Tetracycline                                                                0,5  0,5 20   20                                     ATCC 11 303                                                                   Escherichia Coli Exp. TO.sub.26 B.sub.6                                                                1    1   100  100                                    Escherichia Coli Resistant Gentamicine,                                                                1    1   100  100                                    Tobramycine R 55 123 D                                                        Klebsiella pneumoniae Exp. 52 145                                                                      0,2  0,2 40   40                                     Klebsiella pneumoniae 2 536 Resistant                                                                  10   10  >100 >100                                   Gentamicine                                                                   Proteus mirabilis (indol-) A 235                                                                       0,5  0,5 10   20                                     Proteus vulgaris (indol+) A 232                                                                        0,5  1   >100 >100                                   Salmonella typhimurium 420                                                                             2    2   100  100                                    Providencia Du 48        5    5   >100 >100                                   Pseudomonas 3 935 Exp. SG                                                                              40   40  >100 >100                                   Serratia Resistant Gentamicine 2 532                                                                   2    5   >100 >100                                   Enterobacter cloacae 681 40   40  100  100                                    __________________________________________________________________________    PRODUCT OF EXAMPLE 28                                                                                        M.I.C. in μg/ml                                     Strain                 24 H 48 H                                      __________________________________________________________________________            Staphylococcus aureus ATCC 6 538 Pen-Sensible                                                        0.5  0.5                                               Staphylococcus aureus UC 1 128 Pen-Resistant                                                         1    1                                                 Staphylococcus aureus exp. No 54 146                                                                 1    1                                                 Streptococcus pyogenes A 561                                                                         0.02 0.02                                              Streptococcus faecalis 5 432                                                                         5    5                                                 Streptococcus faecalis 99 F 74                                                                       10   40                                                Bacillus substilis ATCC 6 633                                                                        0.5  1                                                 Escherichia Coli Sensible Tetracycline                                                               0.05 0.1                                               ATCC 9 637                                                                    Escherichia Coli Resistant Tetracycline                                                              0.1  0.1                                               ATCC 11 303                                                                   Escherichia Coli Exp. TO.sub.26 B.sub.6                                                              0.05 0.1                                               Escherichia Coli Resistant Gentamicine                                                               0.2  0.2                                               Tobramycine R 55 123 D                                                        Klebsiella pneumoniae Exp. 52 145                                                                    0.05 0.1                                               Klebsiella pneumoniae 2 536 Resistant                                                                0.2  0.5                                               Gentamicine                                                                   Proteus mirabilis (indol-) A 235                                                                     0.1  0.2                                               Salmonella typhimurium 420                                                                           0.1  0.1                                               Enterobacter cloacae 681                                                                             5    10                                                Providencia Du 48      2    2                                                 Serratia Resistant Gentamicine 2 532                                                                 5    10                                        __________________________________________________________________________

B. Experimental Infection with Escherichia Coli (T) O₂₆ B₆

The product of Example 4 was studied for its activity against anexperimental infection of Escherichia Coli in groups of 10 male miceweighing about 21.5 g. The mice receivedan intraperitoneal injection of0.5 ml of a 24 hour old culture in a nutritive media of Escherichia Coli(T) O₂₆ B₆ of the Pasteur Institute dilute 1/6 with distilled water. Thetest product was administered subcutaneously or orally 1 hour and 5 and24 hours after the infection and the number of dead was determined after8 days. The results are reported in the following Table.

    ______________________________________                                                                  mice alive                                                    Mortality After after 8                                             Dose in mg  211/2 H  281/2 H  32 H  days                                      ______________________________________                                        Controls    9                 1      0/10                                     0.05 subcutaneously                 10/10                                     0.1 subcutaneously                  10/10                                     0.25 subcutaneously                 10/10                                     0.1 subcutaneously                  10/10                                     0.1 orally           1               9/10                                     0.25 orally                         10/10                                     0.5 orally                          10/10                                     ______________________________________                                    

The procedure was repeated with mice weighing about 22.5 g with anintraperitoneal injection of a 24 hour old culture of Escherichia Coli(T) O₂₆ B₆ diluted 1/5.5 with distilled water with the product ofExample 4. The results are reported in the following Table.

    __________________________________________________________________________                    MORTALITY AFTER                                                         Method of                                                                           7 H                                                                              9 H                                                                              22 H                                                                             23 H  25 H                                                                              26 H     46 H     Mice alive on the        Dose in mg                                                                              Treatment                                                                           45 35 30 50 24 H                                                                             50  30 31 H                                                                             32 H                                                                             30 70 H                                                                             4 j                                                                              8th                      __________________________________________________________________________                                                         Day                      Controls  S.C.  1  1  8                              0/10                     3 × 0,5 ml of eau                                                       0,015     S.C.        2  2                  2        4/10                     0,025     S.C.                 1   1              2  6/10                     0,05      S.C.                                       10/10                    0,05      OS          4     1         1  2           2/10                     0,1       OS       1        1            2     1     5/10                     0,25      OS                                         10/10                    __________________________________________________________________________

c. Experimental Infection with Salmonella Typhimurium

Groups of 10 mice with an average weight of 20.5 g wereintraperitoneally injected with 0.5 ml of a 24 hour old culture in oxoidbouillon of Salmonella typhimurium 5210 diluted 1/75 with distilledwater. The product of Example 7 was administered subcutaneously one hourand 4,8,24 and 32 hours after the infection and the results, determinedas before, are reported in the following Table.

    __________________________________________________________________________    MORTALITY AFTER                                  Mice alive on                Dose in mg                                                                          21 H 30                                                                            25 H                                                                             28 H                                                                             29 H                                                                             45 H 30                                                                            56 H                                                                             70 H                                                                             4 J                                                                              5 J                                                                              6 J                                                                              7 J                                                                              8 J                                                                              9 J                                                                              the 10th                     __________________________________________________________________________                                                     day                          Controls                                                                            1    1  1  1  1    1  4                    0/10                         0,1                                  3  3  1     3/10                         0,25                                    2  1  2  5/10                         0,5                               1  1  1        7/10                         1                                          1     9/10                         2                                                10/10                        __________________________________________________________________________

D. Experimental infection of Proteus Mirabilis

Groups of 10 mice with an average weight of 22 g receivedintraperitonally 0.5 ml of a 24 hour old culture in oxoid bouillon ofProteus Mirabilis A 235 diluted 1/4 with distilled water. The mice thenreceived subcutaneously the compound of Example 11 or 15 one hour and 5and 24 hours after the infection and the results are reported in thefollowing Table.

    __________________________________________________________________________    Product    MORTALITY AFTER                                                    Dose in                                                                             of   6 H                                                                              21 H                                                                             22 H                                                                             23 H                                                                              23 H                                                                             23 H                                                                             24 H      31 H           Mice alive on          mg    Examples                                                                           55 30 15 10  30 45 20 28 H                                                                             29 H                                                                              45 46 H                                                                             53 H                                                                             70 H                                                                             94 H                                                                             the 8th                __________________________________________________________________________                                                           day                    Controls   1  9                                        0/10                   0.025 11      3  1            1  2         2  1        0/10                   0.05  11         1            1         1        2     5/10                   0.1   11                         1         2           7/10                   0.25  11                                               10/10                  0.5   11                                               10/10                  1     11                                               10/10                  0.5   15   1  9                                        0/10                   1     15      5     1   1        2               1     0/10                   __________________________________________________________________________

E. Acute Toxicity

The acute toxicity of the product of Example 7 was determined on groupsof 8 to 10 female mice of Swiss CD1 Specific Pathogon Free Strain (stockCH.River-France), with an average weight of 19 to 21 g, for each dose ofthe compound.

The animals had injection in the caudal vein in a variable volume at arate of 1 ml/mn of the product of Example 7 dissolved at a concentrationof 100 mg/ml in sterile apyrogen distilled water. The animals wereobserved for 4 days with food and drinking water "ad libitum" at atemperature of 21°±1° C., constant humidity and area with excess airpressure. The results are reported in the following Table.

    ______________________________________                                        Dose in mg    Total Mortality                                                 ______________________________________                                         500          0/10                                                            1000          0/10                                                            2000          0/8                                                             ______________________________________                                    

The lethal dose of the compound of Example 7 is therefore above 2000mg/kg.

Symptomatology

No symptoms of intoxication were noted following the injections norduring the observation period.

Various modifications of the products and processes of the invention maybe made without departing from the spirit or scope thereof and it shouldbe understood that the invention is to be limited only as defined in theappended claims.

We claim:
 1. The syn isomer of ethyl2-(2-amino-4-thiazolyl)-2-hydroxyimino-acetate substantially free of theanti isomer.